Compounds, pharmaceutical compositions, and methods for inhibiting cyclin-dependent kinases

ABSTRACT

Pharmaceutical compositions containing effective amounts of CDK-inhibiting diaminothiazole compounds of the following formula (where R 1  and R 2  are as defined in the specification) or their salts, or prodrugs or active metabolites of such compounds or salts, are useful for treating disorders and diseases such as cancer:  
                 
 
     In preferred embodiments, R 1  and R 2  are independently unsubstituted or substituted carbocyclic or heterocyclic aryl ring structures. Compounds where R 2  is ortho-substituted aryl are especially potent inhibitors of CDKs such as CDK4.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This regular application claims priority to U.S. ProvisionalApplication No. 60/063,634, filed Oct. 27, 1997, and U.S. ProvisionalApplication No. 60/063,666, filed Oct. 28, 1997.

FIELD OF THE INVENTION

[0002] This invention is directed to pharmaceutical compositionscontaining aminothiazole compounds for inhibiting cyclin-dependentkinases (CDKs), such as CDK1, CDK2, CDK4, and CDK6. The invention isalso directed to the therapeutic or prophylactic use of pharmaceuticalcompositions containing such compounds and to methods of treatingmalignancies and other disorders by administering effective amounts ofsuch compounds.

BACKGROUND OF THE INVENTION

[0003] Uncontrolled cell proliferation is the insignia of cancer. Cellproliferation in response to various stimuli is manifested by aderegulation of the cell division cycle, the process by which cellsmultiply and divide. Tumor cells typically have damage to the genes thatdirectly or indirectly regulate progression through the cell divisioncycle.

[0004] CDKs constitute a class of enzymes that play critical roles inregulating the transitions between different phases of the cell cycle,such as the progression from a quiescent stage in G₁ (the gap betweenmitosis and the onset of DNA replication for a new round of celldivision) to S (the period of active DNA synthesis), or the progressionfrom G₂ to M phase, in which active mitosis and cell-division occur.See, e.g., the articles compiled in Science, vol. 274 (1996), pp.1643-1677; and Ann. Rev. Cell Dev. Biol., vol. 13 (1997), pp. 261-291.CDK complexes are formed through association of a regulatory cyclinsubunit (e.g., cyclin A, B1, B2, D1, D2, D3, and E) and a catalytickinase subunit (e.g., cdc2 (CDK1), CDK2, CDK4, CDK5, and CDK6). As thename implies, the CDKs display an absolute dependence on the cyclinsubunit in order to phosphorylate their target substrates, and differentkinase/cyclin pairs function to regulate progression through specificportions of the cell cycle.

[0005] The D cyclins are sensitive to extracellular growth signals andbecome activated in response to mitogens during the G₁ phase of the cellcycle. CDK4/cyclin D plays an important role in cell cycle progressionby phosphorylating, and thereby inactivating, the retinoblastoma protein(Rb). Hypophosphorylated Rb binds to a family of transcriptionalregulators, but upon hyperphosphorylation of Rb by CDK4/cyclin D, thesetranscription factors are released to activate genes whose products areresponsible for S phase progression. Rb phosphorylation and inactivationby CDK4/cyclin D permit passage of the cell beyond the restriction pointof the G₁ phase, whereupon sensitivity to extracellular growth orinhibitory signals is lost and the cell is committed to cell division.During late G₁, Rb is also phosphorylated and inactivated by CDK2/cyclinE, and recent evidence indicates that CDK2/cyclin E can also regulateprogression into S phase through a parallel pathway that is independentof Rb phosphorylation (see Lukas et al., “Cyclin E-induced S PhaseWithout Activation of the pRb/E2F Pathway,” Genes and Dev., vol. 11(1997), pp. 1479-1492).

[0006] The progression from G₁ to S phase, accomplished by the action ofCDK4/cyclin D and CDK2/cyclin E, is subject to a variety of growthregulatory mechanisms, both negative and positive. Growth stimuli, suchas mitogens, cause increased synthesis of cyclin D1 and thus increasedfunctional CDK4. By contrast, cell growth can be “reined in,” inresponse to DNA damage or negative growth stimuli, by the induction ofendogenous inhibitory proteins. These naturally occurring proteininhibitors include p21^(WAF1/CIP1), p27^(KIP1), and the p16^(INK4)family, the latter of which inhibit CDK4 exclusively (see Harper,“Cyclin Dependent Kinase Inhibitors,” Cancer Surv., vol. 29 (1997), pp.91-107). Aberrations in this control system, particularly those thataffect the function of CDK4 and CDK2, are implicated in the advancementof cells to the highly proliferative state characteristic ofmalignancies, such as familial melanomas, esophageal carcinomas, andpancreatic cancers (see, e.g., Hall and Peters, “Genetic Alterations ofCyclins, Cyclin-Dependent Kinases, and CDK Inhibitors in Human Cancer,”Adv. Cancer Res., vol. 68 (1996), pp.67-108; and Kamb et al., “A CellCycle Regulator Potentially Involved in Genesis of Many Tumor Types,”Science, vol. 264 (1994), pp. 436-440). Over-expression of cyclin D1 islinked to esophageal, breast, and squamous cell carcinomas (see, e.g.,DelSal et al., “Cell Cycle and Cancer: Critical Events at the G₁Restriction Point,” Critical Rev. Oncogenesis, vol. 71 (1996), pp.127-142). Genes encoding the CDK4-specific inhibitors of the p16 familyfrequently have deletions and mutations in familial melanoma, gliomas,leukemias, sarcomas, and pancreatic, non-small cell lung, and head andneck carcinomas (see Nobori et al., “Deletions of the Cyclin-DependentKinase-4 Inhibitor Gene in Multiple Human Cancers,” Nature, vol. 368(1994), pp. 753-756). Amplification and/or overexpression of cyclin Ehas also been observed in a wide variety of solid tumors, and elevatedcyclin E levels have been correlated with poor prognosis. In addition,the cellular levels of the CDK inhibitor p27, which acts as both asubstrate and inhibitor of CDK2/cyclin E, are abnormally low in breast,colon, and prostate cancers, and the expression levels of p27 areinversely correlated with the stage of disease (see Loda et al.,“Increased Proteasome-dependent Degradation of the Cyclin-DependentKinase Inhibitor p27 in Aggressive Colorectal Carcinomas,” NatureMedicine, vol. 3 (1997), pp. 231-234). The p21 proteins also appear totransmit the p53 tumor-suppression signal to the CDKs; thus, themutation of p53 in approximately 50% of all human cancers may indirectlyresult in deregulation of CDK activity.

[0007] The emerging data provide strong validation for the use ofcompounds inhibiting CDKs, and CDK4 and CDK2 in particular, asanti-proliferative therapeutic agents. Certain biomolecules have beenproposed for this purpose. For example, U.S. Pat. No. 5,621,082 to Xionget al. discloses nucleic acid encoding an inhibitor of CDK6, andEuropean, Patent Publication No. 0 666 270 A2 describes peptides andpeptide mimetics that act as inhibitors of CDK1 and CDK2. Several smallmolecules have been identified as CDK inhibitors (for a recent review,see Webster, “The Therapeutic Potential of Targeting the Cell Cycle,”Exp. Opin. Invest. Drugs, vol. 7 (1998), pp. 865-887). The flavoneflavopiridol displays modest selectivity for inhibition of CDKs overother kinases, but inhibits CDK4, CDK2, and CDK1 equipotently, withIC₅₀s in the 0.1-0.3 μM range. Flavopiridol is currently in Phase IIclinical trials as an oncology chemotherapeutic (Sedlacek et al.,“Flavopiridol (L86-8275; NSC 649890), A New Kinase Inhibitor for TumorTherapy,” Int. J. Oncol., vol. 9 (1996), pp. 1143-1168). Analogs offlavopiridol are the subject of other publications, for example, U.S.Pat. No. 5,733,920 to Mansuri et al. (International Publication No. WO97/16447) and International Publication Nos. WO 97/42949, and WO98/17662. Results with purine-based derivatives are described in Schowet al., Bioorg. Med Chem. Lett., vol. 7 (1997), pp. 2697-2702; Grant etal., Proc. Amer. Assoc. Cancer Res,. vol. 39 (1998), Abst. 1207;Legravend et al., Bioorg. Med. Chem. Lett., vol. 8 (1998), pp. 793-798;Gray et al., Science, vol. 281 (1998), pp. 533-538; and Furet et al.,216th ACS Natl. Mtg. (Aug. 23-27, 1998, Boston), Abst MED1-218. Inaddition, the following publications disclose certain pyrimidines thatinhibit cyclin-dependent kinases and growth-factor mediated kinases:International Publication No. WO 98/33798; Ruetz et al., Proc. Amer.Assoc. Cancer Res,. vol. 39 (1998), Abst. 3796; and Meyer et al., Proc.Amer. Assoc. Cancer Res., vol. 39 (1998), Abst. 3794.

[0008] There is still a need, however, for small-molecule compounds thatmay be readily synthesized and are potent inhibitors of one or more CDKsor CDK/cyclin complexes. Because CDK4 may serve as a general activatorof cell division in most cells, and because complexes of CDK4/cyclin Dand CDK2/cyclin E govern the early G₁ phase of the cell cycle, there isa need for effective and specific inhibitors of CDK4 and/or CDK2 fortreating one or more types of tumors.

SUMMARY OF THE INVENTION

[0009] Accordingly, one object of the invention is to attain compoundsand drug compositions that inhibit the activity of one or more CDKs,such as CDK2, CDK4, and/or CDK6, or cyclin complexes thereof. A furtherobject is to provide an effective method of treating cancer indicationsthrough CDK inhibition, preferably through inhibition of CDK4 orCDK4/D-type cyclin complexes and/or CDK2 or CDK2/E-type cyclincomplexes. Another object is to achieve pharmaceutical compositionscontaining compounds effective to block the transition of cancer cellsinto their proliferative phase. These and other objects and advantagesof the invention, which will become apparent in light of the detaileddescription below, are achieved through cell-cycle control agents of theinvention described below.

[0010] In one general aspect, the invention relates to pharmaceuticalcompositions comprising:

[0011] (a) a cell-cycle control agent selected from:

[0012] (i) compounds of the Formula 1:

[0013]  wherein:

[0014] R¹ is a substituted or unsubstituted group selected from:C₁₋₆-alkyl (e.g., methyl, ethyl, propyl, isopropyl, butyl, isobutyl,sec-butyl, or tert-butyl); C₁₋₆-alkenyl; C₁₋₆-alkynyl; C₁₋₆-alkoxyl;C₁₋₆-alcohol; carbocyclic or heterocyclic cycloalkyl, which may bemonocyclic or fused or non-fused polycyclic (e.g., cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl) or heterocycloalkyl, which may bemonocyclic or fused or non-fused polycyclic (e.g., pyrrolidinyl,piperidinyl, morpholinyl); carbocyclic or heterocyclic, monocyclic orfused or non-fused polycyclic aryl (e.g., phenyl, naphthyl, pyrrolyl,indolyl, furanyl, thiophenyl, imidazolyl, oxazolyl, isoxazolyl,thiazolyl, triazolyl, tetrazolyl, pyrazolyl, pyridinyl, quinolinyl,isoquinolinyl, acridinyl, pyrazinyl, pyridazinyl, pyrimidinyl,benzimidazolyl, benzothiophenyl, or benzofuranyl); carbonyl (e.g.,carboxyl, ester, aldehyde, or ketone); ether; (C₁₋₆-alkyl)-carbonyl;(C₁₋₆-alkyl)-aryl; (C₁₋₆-alkyl)-cycloalkyl; (C₁₋₆-alkyl)-(C₁₋₆-alkoxyl);aryl-(C₁₋₆-alkoxyl); thioether (e.g., aryl-S-aryl, cycloalkyl-S-aryl,cycloalkyl-S-cycloalkyl, or dialkyl sulfide); thiol; and sulfonyl; and

[0015] R² is a substituted or unsubstituted: carbocyclic orheterocyclic, monocyclic or fused or non-fused polycyclic, ringstructure;

[0016] where each optional substituent for R¹ and R² is independently ahalogen (e.g., chloro, iodo, bromo, or fluoro); oxygen (═O); haloalkyl(e.g., trifluoromethyl); C₁₋₆-alkyl; C₁₋₆-alkenyl; C₁₋₆-alkynyl;hydroxyl; C₁₋₆-alkoxyl; carbocyclic cycloalkyl, which may be monocyclicor fused or non-fused polycyclic (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl), or a heterocycloalkyl, which may bemonocyclic or fused or non-fused polycyclic (e.g., pyrrolidinyl,piperidinyl, piperazinyl, morpholinyl, or thiazinyl); carbocyclic orheterocyclic, monocyclic or fused or non-fused polycyclic aryl (e.g.,phenyl, naphthyl, pyrrolyl, indolyl, furanyl, thiophenyl, imidazolyl,oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, pyrazolyl,pyridinyl, quinolinyl, isoquinolinyl, acridinyl, pyrazinyl, pyridazinyl,pyrimidinyl, benzimidazolyl, benzothiophenyl, or benzofuranyl); amino(primary, secondary, or tertiary); nitro; thiol; thioether; imine;cyano; amido; phosphonato; phosphine; carboxyl; thiocarbonyl; sulfonyl;sulfonamide; ketone; aldehyde; or ester;

[0017] (ii) pharmaceutically acceptable salts of compounds of theFormula I; and

[0018] (iii) prodrugs and pharmaceutically active metabolites ofcompounds of the Formula I or pharmaceutically acceptable salts thereof;and

[0019] (b) a pharmaceutically acceptable carrier.

[0020] In a further general aspect, the invention relates topharmaceutical compositions comprising:

[0021] (a) a cell-cycle control agent selected from:

[0022] (i) compounds of the Formula I:

[0023]  wherein:

[0024] R² is a substituted or unsubstituted: carbocyclic orheterocyclic, monocyclic or fused or non-fused polycyclic, ringstructure; where each optional substituent for R² is independently ahalogen (e.g., chloro, iodo, bromo, or fluoro); oxygen (═O); haloalkyl(e.g., trifluoromethyl); C₁₋₆-alkyl; C₁₋₆-alkenyl; C₁₋₆-alkynyl;hydroxyl; C₁₋₆-alkoxyl; carbocyclic cycloalkyl, which may be monocyclicor fused or non-fused polycyclic (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl), or a heterocycloalkyl, which may bemonocyclic or fused or non-fused polycyclic (e.g., pyrrolidinyl,piperidinyl, piperazinyl, morpholinyl, or thiazinyl); carbocyclic orheterocyclic, monocyclic or fused or non-fused polycyclic aryl (e.g.,phenyl, naphthyl, pyrrolyl, indolyl, furanyl, thiophenyl, imidazolyl,oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, pyrazolyl,pyridinyl, quinolinyl, isoquinolinyl, acridinyl, pyrazinyl, pyridazinyl,pyrimidinyl, benzimidazolyl, benzothiophenyl, or benzofuranyl); amino(primary, secondary, or tertiary); nitro; thiol; thioether; imine;cyano; amido; phosphonato; phosphine; carboxyl; thiocarbonyl; sulfonyl;sulfonamide; ketone; aldehyde; or ester;

[0025] (ii) pharmaceutically acceptable salts of compounds of theFormula I; and

[0026] (iii) prodrugs and pharmaceutically active metabolites ofcompounds of the Formula I or pharmaceutically acceptable salts thereof;and

[0027] (b) a pharmaceutically acceptable carrier.

[0028] Such compositions are useful as inhibitors of mammalianCDK/cyclin complexes, insect CDK, or fungal CDK complexes. Suchcompositions are also useful for controlling proliferation,differentiation, and/or apoptosis. Thus, in one general aspect theinvention is directed to pharmaceutical compositions containingpharmaceutically effective amounts of cell-cycle control agents.

[0029] In a preferred embodiment, the invention is directed to potentcell-cycle control agents where R² in Formula I is an ortho-substitutedaryl ring structure (e.g., o-substituted phenyl). Particularly preferredamong such agents are those in which R² is an o-disubstituted phenyl.

[0030] The invention further relates to methods of using cell-cyclecontrol agents for treating diseases or disorders mediated by CDKinhibition, especially those mediated by CDK4 and/or CDK2 inhibition.More particularly, the invention is directed to methods of treatingmalignancies or cancer-type disorders by administering a pharmaceuticalcomposition comprising a cell-cycle control agent. Additionally, theinvention relates to the use of cell-cycle control agents to prevent ortreat mycotic infections.

[0031] Other aspects, advantages, and preferred features of theinvention will become apparent from the detailed description below.

DETAILED DESCRIPTION AND PREFERRED EMBODIMENTS OF THE INVENTION

[0032] In one general embodiment, the invention relates topharmaceutical compositions each comprising:

[0033] (a) an amount of a cell-cycle control agent effective to inhibita CDK, the cell-cycle control agent being:

[0034] (i) a compound of the Formula I:

[0035]  wherein:

[0036] R¹ is a substituted or unsubstituted group selected from:C₁₋₆-alkyl; C₁₋₆-alkenyl; C₁₋₆-alkynyl; C₁₋₆-alkoxyl; carbocylic orheterocyclic, monocyclic or fused or non-fused polycyclic, cycloalkyl;carbocyclic or heterocyclic, monocyclic or fused or non-fusedpolycyclic, aryl; carbonyl; ether; (C₁₋₆-alkyl)-carbonyl;(C₁₋₆-alkyl)-aryl; (C₁₋₆-alkyl)-cycloalkyl; (C₁₋₆-alkyl)-(C₁₋₆-alkoxyl);aryl-(C₁₋₆-alkoxyl); thioether; thiol; and sulfonyl; and

[0037] R² is a substituted or unsubstituted, carbocyclic orheterocyclic, monocyclic or fused or non-fused polycyclic, ringstructure;

[0038] where each optional substituent for R¹ and R² is independently ahalogen; haloalkyl; C₁₋₆-alkyl; C₁₋₆-alkenyl; C₁₋₆-alkynyl; hydroxyl;C₁₋₆ alkoxyl; amino; nitro; thiol; thioether; imine; cyano; amido;phosphonato; phosphine; carboxyl; thiocarbonyl; sulfonyl; sulfonamide;ketone; aldehyde; or ester;

[0039] (ii) a pharmaceutically acceptable salt of a compound of theFormula I; or

[0040] (iii) a prodrug or pharmaceutically active metabolite of acompound of the Formula I or a pharmaceutically acceptable salt thereof;and

[0041] (b) a pharmaceutically acceptable carrier.

[0042] In another general embodiment, each optional substituent for R¹and R² may be independently selected from, in addition to theabove-identified groups, the following groups: oxygen; carbocyclic orheterocyclic, monocyclic or fused or non-fused polycyclic, cycloalkyl;and carbocyclic or heterocyclic, monocyclic or fused or non-fusedpolycyclic, aryl. Such substituents may optionally be furthersubstituted with a substituent selected from such groups.

[0043] Examples for the moiety R¹ include substituted or unsubstitutedaryl and alkyl, such as phenyl, pyridyl, benzimidazole, benzyl, andC₁₋₆-alkyl. In a preferred embodiment, these groups have one or moresubstituents selected from: halogen; oxygen; haloalkyl; C₁₋₆-alkyl;cycloalkyl; heterocycloalkyl; aryl; hydroxyl; C₁₋₆ alkoxyl; amino; nitrothioether; cyano; amido; carboxyl; sulfonamido; ketone; aldehyde; andester.

[0044] Other preferred moieties for R¹ are phenyl groups substituted byan alkylamine or pyridine group having optional substituents selectedfrom the group described in the above paragraph for R¹. The alkylaminesubstitutent may be a 5- to 7-membered heterocycloalkyl optionallycontaining, in addition to the nitrogen ring atom, one or moreheteroatoms selected from N, O and S.

[0045] Examples of such preferred R¹ groups include phenyl substitutedin the para position with a heterocycloalkyl, for example piperidinyl,piperazinyl, thiazinyl, or morpholinyl, or a pyridyl group. Thefollowing are examples of preferred R¹ groups:

[0046] Other particularly preferred R¹ groups include phenyl groupssubstituted with carbonyl or sulfonamide moieties, wherein the carbonylcarbon and sulfonamide nitrogen are optionally further substituted. Thefollowing are examples of preferred R¹ groups:

[0047] where R³ is selected from C₁-C₆ alkyl, C₁-C₆ alkoxy, aryl,aryloxy, and amine.

[0048] Other preferred examples for the moiety R¹ include substituted orunsubstituted phenyl, alkylbenzyl, alkyl, benzyl carboxyl ester,benzyloxyphenyl, dimethylaminophenyl, pyridinyl, phenethyl,alkylcarboxyl, alkylpiperidinyl, phenylamino, cyclohexyl,benzylcarboxylalkyl, benzylnitro, phenyl-alkoxyl, ethyl benzoate, benzylcarboxyl, alkylbenzoimidazole, benzoimidazole, benzyldimethylamino,pyridinyl-sulfanyl, cyanobenzyl, and phenyl sulfamyl.

[0049] In preferred embodiments, R² in Formula I is a bulky group suchas a substituted or unsubstituted carbocyclic or heterocyclic monocycle,or a substituted or unsubstituted fused or non-fused carbocyclic orheterocyclic polycycle. More preferably, R² is a substituted (carbo orpoly)-(monocycle or polycycle); even more preferably, R² is such acyclic ring structure bearing a substituent at the position adjacent orvicinal to the point of attachment (to the core structure).

[0050] For example, preferred species for R² include anortho-substituted aromatic ring structure such as o-substituted phenylor thienyl, or a 1,2-substituted cycloalkyl or cycloalkenyl ringstructure such as 2-substituted cyclopent-1-enyl. Particularly preferredexamples for the moiety R² include substituted or unsubstituted:o-halophenyl (e.g., o-fluorophenyl, o-chlorophenyl, o-iodophenyl, oro-bromophenyl), o-nitrophenyl, o-aminophenyl, o-C₁₋₆-alkylphenyl,o-C₁₋₆-alkoxyphenyl (e.g., o-methoxyphenyl or o-ethoxyphenyl),o-C₁₋₆-alkoxybenzothiophenyl, o-methylthiophenyl, benzonitrile, andcarboxybenzyl. Particularly preferred examples for the moiety R² alsoinclude ortho-disubstituted aryls, for example, 2,6-dihalophenyl (e.g.,2,6-difluorophenyl) and 2-halo-6-trifluoromethylphenyl (e.g.,2-fluoro-6-trifluoromethylphenyl). Compounds of the Formula I where R²is a 1,2-substituted cyclic ring structure, optionally having one ormore additional substituents, such as an ortho-substituted aryl havinganother substituent at the para position, have been surprisingly foundto be potent CDK inhibitors.

[0051] Particularly preferred examples of compounds of Formula Iinclude:

[0052] Other particularly preferred examples of compounds of Formula Iinclude:

[0053] Pharmaceutical compositions according to the invention may,alternatively or in addition to a compound of the Formula I, comprise asan active ingredient a pharmaceutically acceptable salt of a compound ofthe Formula I, or a prodrug or pharmaceutically active metabolite ofsuch a compound or salt. Such compounds, salts, prodrugs, andmetabolites are sometimes referred to herein collectively as “cell-cyclecontrol agents.”

[0054] Compositions in accordance with the invention inhibit the kinaseactivity of CDK/cyclin complexes, such as those active in the G₀ or G₁stage of the cell cycle, e.g., CDK2, CDK4, and/or CDK6 complexes.Preferred compositions of the invention contain cell-cycle controlagents having an inhibition constant against CDK4 or a CDK4/D-typecyclin complex of about 1 μM or less, more preferably of about 500 nM orless, even more preferably of about 200 nM or less, and most preferablyof about 100 nM or less. Especially preferred compounds of the inventioninclude those having a CDK4/cyclin D3 inhibition constant (K_(i)CDK4/D3) of about 100 nM or less. Other preferred compositions of theinvention contain cell-cycle control agents having an inhibitionconstant against CDK2 or a CDK2/E-type cyclin complex of about 1 μM orless, more preferably of about 500 nM or less, even more preferably ofabout 200 nM or less, and most preferably of about 100 nM or less.

[0055] Certain compounds of the Formula I may exist in variousstereoisomeric or tautomeric forms. The present invention encompassesall such CDK-inhibiting compounds, including active compounds in theform of essentially pure enantiomers, racemic mixtures, and tautomers.

[0056] The term “pharmaceutically acceptable” means pharmacologicallyacceptable and substantially non-toxic to the subject being administeredthe cell-cycle control agent. Pharmaceutically acceptable salts includeconventional acid-addition salts or base-addition salts formed fromsuitable non-toxic organic or inorganic acids or inorganic bases.Exemplary acid-addition salts include those derived from inorganic acidssuch as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuricacid, sulfamic acid, phosphoric acid, and nitric acid, and those derivedfrom organic acids such as p-toluenesulfonic acid, methanesulfonic acid,ethane-disulfonic acid, isethionic acid, oxalic acid,p-bromophenylsulfonic acid, carbonic acid, succinic acid, citric acid,benzoic acid, 2-acetoxybenzoic acid, acetic acid, phenylacetic acid,propionic acid, glycolic acid, stearic acid, lactic acid, malic acid,tartaric acid, ascorbic acid, maleic acid, hydroxymaleic acid, glutamicacid, salicylic acid, sulfanilic acid, and fumaric acid. Exemplarybase-addition salts include those derived from ammonium hydroxides(e.g., a quaternary ammonium hydroxide such as tetramethylammoniumhydroxide), those derived from inorganic bases such as alkali oralkaline earth-metal (e.g., sodium, potassium, lithium, calcium, ormagnesium) hydroxides, and those derived from organic bases such ascarbonates, bicarbonates, amines, benzylamines, piperidines, andpyrrolidines.

[0057] The term “prodrug” refers to a metabolic precursor of a compoundof the Formula I (or a salt thereof) that is pharmaceuticallyacceptable. A prodrug may be inactive when administered to a subject butis converted in vivo to an active compound of the Formula I. The term“active metabolite” refers to a metabolic product of a compound of theFormula I that is pharmaceutically acceptable and effective. Prodrugsand active metabolites of compounds of the Formula I may be determinedusing techniques known in the art.

[0058] Cell-cycle control agents in accordance with the invention areuseful as pharmaceuticals for treating proliferative disorders inmammals, especially humans, marked by unwanted proliferation ofendogenous tissue. Compounds of the Formula I may be used for treatingsubjects having a disorder associated with excessive cell proliferation,e.g., cancers, psoriasis, immunological disorders involving undesiredproliferation of leukocytes, and restenosis and other smooth-muscledisorders. Furthermore, such compounds may be used to preventde-differentiation of post-mitotic tissue and/or cells.

[0059] Pharmaceutical compositions or preparations of the inventioncomprise a pharmaceutically acceptable carrier and an effective amountof at least one cell-cycle control agent. The term “effective amount”means an amount that significantly inhibits proliferation and/orprevents de-differentiation of a eukaryotic cell, e.g., a mammalian,insect, plant, or fungal cell, and is effective for the indicatedutility, e.g., specific therapeutic treatment.

[0060] The specific dosage amount of a cell-cycle control agent beingadministered to obtain therapeutic or inhibitory effects, of course, maybe determined in a manner known in the art according to the particularcircumstances surrounding the case, including, e.g., the specific agentbeing administered, the route of administration, the condition beingtreated, and the subject or host being treated. An exemplary total dailydose of a cell-cycle control agent, which may be administered in singleor multiple doses, contains a dosage level of from about 0.01 mg/kg bodyweight to about 50 mg/kg body weight.

[0061] The cell-cycle control agents of the invention may beadministered by any of a variety of suitable routes, such as orally,rectally, transdermally, subcutaneously, intravenously, intramuscularly,or intranasally. The cell-cycle control agents are preferably formulatedinto compositions suitable for the desired routes before beingadministered.

[0062] A pharmaceutical composition or preparation according to theinvention comprises an effective amount of a cell-cycle control agentand a pharmaceutically acceptable carrier, such as a diluent orexcipient for the agent. When the carrier serves as a diluent, it may bea solid, semi-solid, or liquid material acting as a vehicle, excipient,or medium for the active ingredient(s). Compositions according to theinvention may be made by admixing the active ingredient(s) with acarrier, or diluting it with a carrier, or enclosing or encapsulating itwithin a carrier, which may be in the form of a capsule, sachet, papercontainer, or the like. Exemplary ingredients, in addition to one ormore cell-cycle control agents and any other active ingredients, includeAvicel (microcrystalline cellulose), starch, lactose, calcium sulfatedihydrate, terra alba, sucrose, talc, gelatin, agar, pectin, acacia,magnesium stearate, stearic acid, peanut oil, olive oil, glycerylmonostearate, Tween 80 (polysorbate 80), 1,3-butanediol, cocoa butter,beeswax, polyethylene glycol, propylene glycol, sorbitan monostearate,polysorbate 60, 2-octyldodecanol, benzyl alcohol, glycine, sorbic acid,potassium sorbate, disodium hydrogen phosphate, sodium chloride, andwater.

[0063] The compositions may be prepared in any of a variety of formssuitable for the desired mode of administration. For example,pharmaceutical compositions may be prepared in the form of tablets,pills, powders, lozenges, sachets, cachets, elixirs, suspensions,emulsions, solutions, syrups, aerosols (as solids or in liquid media),ointments (e.g., containing up to 10% by weight of a cell-cycle controlagent), soft-gel and hard-gel capsules, suppositories, sterileinjectable solutions, sterile packaged powders, and the like.

[0064] A pharmaceutical composition according to the invention comprisesa cell-cycle control agent and, optionally, one or more other activeingredients, such as a known antiproliferative agent that is compatiblewith the cell-cycle control agent and suitable for the indication beingtreated. In a preferred embodiment, a pharmaceutical composition of theinvention includes an effective amount of a cell-cycle control agent ofthe Formula I as an active ingredient.

[0065] Compounds in accordance with the invention may be prepared inmanners analogous to those specifically described below, with thelettered example prefixes (i.e., A, B, C, D, E, F, G, H, J, K, L, M andN) designating general synthesis schemes.

EXAMPLES Example A(1)(4-Amino-2-phenylamino-thiazol-5-yl)-(3-nitrophenyl)-methanone

[0066]

[0067] Following the procedure of Gewald et al., J. Prakt. Chem., vol.35 (1967), pp. 97-104, sodium (188 mg, 8.20 mmol) was carefullydissolved in methanol (9 mL) at 0° C. and allowed to warm to ambienttemperature. The resultant solution was added portionwise to a mixtureof cyanamide (345 mg, 8.20 mmol) and phenyl isothiocyanate (0.98 mL, 8.2mmol), whereupon heat evolved. 2-Bromo-3′-nitroacetophenone (2.00 g, 8.2mmol) was added, and the resultant mixture stirred overnight at ambienttemperature. The mixture was diluted with water (150 mL). A yellow-brownsolid was filtered off, rinsed with water and a small amount of ether,dried under vacuum, and recrystallized from ethanol to give 2.17 g (52%yield) of the title compound in the form of dark brown crystals, meltingpoint (mp) 186-187° C.

[0068]¹-H NMR (DMSO-d₆): δ 10.95 (1H, s), 8.44 (1H, t, J=1.9 Hz),8.54-8.22 (2H, bs), 8.34 (1H, ddd, J=8.2, 1.9, 0.9 Hz), 8.12 (1H, ddd,J=8.2, 1.9, 0.9 Hz), 7.78 (1H, t, J=8.2 Hz), 7.62 (2H, d, J=7.8 Hz),7.36 (2H, t, J=7.8 Hz), 7.09 (1H, t, J=7.8 Hz).

[0069] ESIMS (MH⁺): 341.

[0070] Anal. Calcd. for C₁₆H₁₂N₄O₃S.EtOH: C, 55.94; H, 4.70; N, 14.50;S, 8.30. Found: C, 55.96; H, 4.73; N, 14.40; S, 8.29.

Example A(2)(4-amino-2-phenylamino-thiazol-5-yl)-(4-nitrophenyl)-methanone

[0071]

[0072] The title compound was prepared in a manner analogous to thatdescribed for Example A(1). Phenyl isothiocyanate and2-bromo-4′-nitro-acetophenone gave, after recrystallization fromethanol, 3.06 g (55% yield) of red-brown crystals, mp 162-164° C.

[0073]¹H NMR (DMSO-d₆): δ 10.91 (1H, s), 8.38 (2H, bs), 8.30 (2H, d,J=8.7 Hz), 7.90 (2H, d, J=8.7 Hz), 7.59 (2H, d, J=7.5 Hz), 7.36 (2H, t,J=7.5 Hz), 7.10 (1H, d, J=7.5 Hz).

[0074] FABMS (MH⁺): 341.

[0075] Anal. Calcd. for C₁₆H₁₂N₄O₃S: C, 56.46; H, 3.55; N, 16.46; S,9.42. Found: C, 56.54; H, 3.54; N, 16.52; S, 9.42.

Example A(3)[4-Amino-2-(pyridin-3-ylamino)-thiazol-5-yl]-phenyl-methanone

[0076]

[0077] The title compound was prepared in a manner similar to thatdescribed for Example A(1). Pyridin-3-yl isothiocyanate and phenacylchloride provided, after recrystallization from ethanol, 4.1 g (75%yield) of yellow crystals, mp 227-229° C.

[0078]¹H NMR (DMSO-d₆): δ 10.95 (1H, s), 8.82(1H, d, J=2.5 Hz), 8.28(1H, dd, J=4.7, 1.2 Hz), 8.23 (2H, bs), 8.12 (1H, ddd, J=8.4, 2.8, 1.6Hz), 7.68 (1H, d, J=6.9 Hz), 7.66 (1H, d, J=7.8 Hz), 7.54-7.44 (3H, m),7.39 (1H, dd, J=8.4, 4.7 Hz).

[0079] HRFABMS: Calcd. for C₁₅H₁₃N₄OS (MH⁺): 297.0810. Found: 297.0815.

[0080] Anal. Calcd. for C₁₅H₁₂N₄OS.EtOH: C, 59.63; H, 5.30; N, 16.36; S.9.36. Found: C, 59.62; H, 5.32; N, 16.43; S, 9.41.

Example A(4) (4-Amino-2-phenylamino-thiazol-5-yl)-pyridin-2-yl-methanone

[0081]

[0082] The title compound was prepared in a manner similar to thatdescribed for Example A(1). Phenyl isothiocyanate and2-(2-bromoacetyl)pyridine (see Menasse et al., Helv. Chim. Acta, vol. 38(1955), pp. 1289-1291; Imuta et al., J. Org. Chem., vol. 45 (1980), pp.3352-3355) gave, after recrystallization from 95% ethanol, 510 mg (71%yield) of brown needles, mp 181.5-183.0° C.

[0083]¹H NMR (DMSO-d₆): δ 10.75 (1H, s), 8.92 (1H, bs), 8.66 (1H, ddd,J=5.1 1.8, 1.2 Hz), 8.22 (1H, bs), 8.13 (1H, dt, J=7.5, 1.2 Hz), 8.01(1H, dt, J=7.5, 1.8 Hz), 7.69 (2H, dd, J=7.5, 1.2 Hz), 7.54 (1H, ddd,J=7.5, 5.1, 1.2 Hz), 7.36 (2H, t, J=7.5 Hz), 7.07 (11, dt, J=7.5, 1.2Hz).

[0084] HRFABMS: Calcd. for C₁₅H₁₃N₄OS (MH⁺): 297.0810. Found: 297.0818.

[0085] Anal. Calcd. for C₁₅H₁₂N₄OS.H₂O: C, 57.31; H, 4.49; N, 17.82; S,10.20. Found: C, 57.31; H, 4.46; N, 17.80; S, 10.14.

Example A(5) Ethyl4-[4-Amino-5-(2-nitro-benzoyl)-thiazol-2-ylamino]-benzoate

[0086]

[0087] The title compound was prepared in a manner analogous to thatused in Example A(1). 4-Carboethoxy-phenyl isothiocyanate and2-bromo-2′-nitro-acetophenone gave, after recrystallization fromethanol, 1.2 g (59% yield) of yellow crystalline powder, mp 262-265° C.

[0088]¹H NMR (DMSO-d₆): δ 11.08 (1H, s), 8.12 (2H, bs), 8.08 (1H, d,J=8.7 Hz), 7.93 (2H, d, J=8.7 Hz), 7.82 (1H, dt, J=7.2, 1.2 Hz),7.77-7.66 (31H, m), 7.73 (1H, d, J=8.7 Hz), 4.28 (2H, q, J=7.2 Hz), 1.30(3H, t, J=7.2 Hz).

[0089] ESIMS (MH⁺): 413.

[0090] Anal. Calcd. for C₁₉H₁₈N₄O₃S: C, 55.33; H, 3.91; N, 13.58; S,7.77. Found: C, 55.22; H, 3.86; N, 13.48; S, 7.67.

Example A(6)[4-Amino-2-(2-methyl-1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(2-nitro-phenyl)-methanone

[0091]

[0092] The title compound was prepared in a similar manner to thatdescribed in Example A(1). 6-Isothiocyanato-2-methyl-1H-benzoimidazole(see Galley et al., German Patent Publication DE 2259220 (1973); C.A.No. 478781 (1973)) and 2-bromo-2′-nitro-acetophenone gave, afterrecrystallization from ethanol, 1.2 g (62% yield) of brown crystals, mp190.0-192.5° C.

[0093]¹H NMR (DMSO-d₆): δ 12.20 (1H, bs), 10.76 (1H, s), 8.10-8.76 (3H,m), 7.76 (1H, t, J=7.0 Hz), 7.70-7.58 (3H, m), 7.40 (1H, d, J=8.4 Hz),7.13 (1H, dd, J=8.4, 1.6 Hz), 2.44 (3H, s).

[0094] FABMS (MH⁺): 395.

[0095] Anal. Calcd. for C₁₈H₁₄N₆O₃S.H₂O: C, 52.42; H, 3.91; N, 20.38; S,7.77. Found: C, 52.29; H, 3.89; N, 20.31; S, 7.86.

Example A(7)[4-Amino-2-(4-iodo-phenylamino)-thiazol-5-yl]-(2-nitro-phenyl)-methanone

[0096]

[0097] The title compound was prepared analogously to Example A(1).4-Iodophenyl isothiocyanate and 2-bromo-2′-nitro-acetophenone provided7.9 g (88% yield) of orange-red powder, mp 182-184° C.

[0098]¹H NMR (DMSO-d₆): δ 10.89 (1H, s), 8.20 (1H, s), 8.50 (1H, d,J=8.7 Hz), 7.80 (1H, dd, J=8.7, 6.2 Hz), 7.72-7.62 (4H, m), 7.41 (2H, d,J=8.7 Hz).

[0099] FABMS (MH⁺): 467.

[0100] Anal. Calcd. for C₁₆H₁₁N₄O₃SI: C, 41.21; H, 2.38; N, 12.02; S,6.88; 1, 27.22. Found: C, 41.34; H, 2.46; N, 12.07; S, 7.02; I, 27.35.

Example A(8)[4-Amino-2-(4-nitro-phenylamino)-thiazol-5-yl]-phenyl-methanone

[0101]

[0102] The title compound was prepared in a manner analogous to thatdescribed for Example A(1). 4-Nitrophenyl isothiocyanate and phenacylchloride furnished 2.5 g (60% yield) of solid, mp 280.0-281.5° C.

[0103]¹H NMR (DMSO-d₆): δ 11.38 (1H, s), 8.30-8.18 (2H, bs), 8.23 (2H,d, J=9.3 Hz), 7.87 (2H, d, J=9.3 Hz), 7.69 (2H, dd, J=7.8, 1.6 Hz),7.56-7.44 (3H, m).

[0104] FABMS (MH⁺): 341.

[0105] Anal. Calcd. for C₁₆H₁₂N₄O₃S: C, 56.46; H, 3.55; N, 16.46; S,9.42. Found: C, 56.40; H, 3.49; N, 16.40; S, 9.41.

Example A(9)[4-Amino-2-(1H-benzoimidazol-6-yl-amino)-thiazol-5-yl]-(2-nitro-phenyl)-methanone

[0106]

[0107] The title compound was prepared in a manner similar to thatdescribed for Example A(1). 6-Isothiocyanato-1H-benzoimidazole (see Boevet al., Pharm. Chem. J. (Engl. Transl.), vol. 24 (1990), pp. 818-822)and 2-bromo-2′-nitro-acetophenone afforded, after recrystallization fromethanol/methanol, 1.5 g (83% yield) of red-brown amorphous powder, mp249-255° C.

[0108]¹H NMR (DMSO-d₆): δ 12.50 (1H, bs), 10.84 (1H, s), 8.20 (1H, s),8.60 (2H, bs), 8.03 (1H, d, J=8.1 Hz), 7.88-7.78 (11H, m), 7.76 (1H, d,J=7.5 Hz), 7.66 (11H, t, J=7.5 Hz), 7.65 (1H, s), 7.55 (1H, d, J=8.7Hz), 7.21 (1H, d, J=8.7 Hz).

[0109] FABMS (MH⁺): 381.

[0110] Anal. Calcd. for C₁₇H₁₂N₆O₃S: C, 53.68; H, 3.18; N, 22.09; S,8.43. Found: C, 53.69; H, 3.14; N, 21.99; S, 8.39.

Example A(10)[4-Amino-2-(4-methoxy-phenylamino)-thiazol-5-yl]-(2-nitro-phenyl)-methanone

[0111]

[0112] The title compound was prepared in a manner similar to thatdescribed for Example A(1). 4-Methoxy-phenyl isothiocyanate and2-bromo-2′-nitro-acetophenone provided, after recrystallization fromaqueous ethanol, 562 mg (43% yield) of brown-red crystals, mp 185-188°C.

[0113]¹H NMR (DMSO-d₆): δ 10.65 (1H, s), 8.25 (2H, bs), 8.20 (1H, d,J=7.5 Hz), 7.77 (1H, t, J=7.5 Hz), 7.66 (1H, t, J=7.5 Hz), 7.62 (1H, d,J=7.5 Hz), 7.41 (2H, d, J=8.7 Hz), 6.92 (2H, d, J=8.7 Hz), 3.72 (3H, s).

[0114] FABMS (M+Na⁺): 393.

[0115] Anal. Calcd. for C₁₇H₁₄N₄O₄S: C, 55.13; H, 3.81; N, 15.13; S,8.66. Found: C, 55.08; H, 3.83; N, 15.11; S, 8.56.

Example A(11)[4-Amino-2-(pyridin-3-ylamino)-thiazol-5-yl]-(2-nitro-phenyl)-methanone

[0116]

[0117] The title compound was prepared as essentially described forExample A(1). Pyridin-3-yl isothiocyanate and2-bromo-2′-nitro-acetophenone afforded, after column chromatography with5% MeOH/CH₂Cl₂, 750 mg (42% yield) of yellow solid, mp 143.5-146.0° C.

[0118]¹H NMR (DMSO-d₆): δ 10.95 (1H, bs), 8.62 (1H, s), 8.19 (1H, dd,J=4.7, 1.3 Hz), 8.08-7.86 (4H, m), 7.76 (1H, td, J=8.3, 0.9 Hz), 7.66(1H, dd, J=8.4, 1.3 Hz), 7.62 (1H, d, J=7.5 Hz), 7.31 (1H, dd, J=8.4,4.7 Hz).

[0119] FABMS (MH⁺): 342.

[0120] Anal. Calcd. for C₁₅H₁₁N₅O₃S.0.5H₂O.0.4 EtOH: C, 51.46; H, 3.94;N, 18.99; S, 8.69. Found: C, 51.86; H, 3.88; N, 19.24; S, 8.88.

Example A(12)4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazole-5-carboxylic Acid MethylEster

[0121]

[0122] The title compound was prepared essentially as described forExample A(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al.,Pharm. Chem. J. (Engl. Transl.). vol. 24 (1990), pp. 818-822) and methylbromoacetate gave in 63% yield a yellow solid, mp 266-267° C.

[0123]¹H NMR (DMSO-d₆): δ 12.37 (1H, bs), 10.52 (1H, s), 8.15 (1H, s),7.92 (1H, s), 7.52 (1H, d, J=8.7 Hz), 7.23 (1H, dd, J=8.7, 1.9 Hz), 6.89(2H, bs), 3.62 (3H, s).

[0124] FABMS (MH⁺): 250.

[0125] Anal. Calcd. for C₁₁H₁₁N₃O₂S.0.15 EtOH: C, 49.87; H, 4.05; N,23.64; S, 10.82. Found: C, 49.94; H, 3.94; N, 23.41; S, 10.79.

Example A(13)[4-Amino-2-(p-tolylamino)-thiazol-5-yl]-(2,4-dimethoxyphenyl)-methanone

[0126]

[0127] The title compound was prepared in a manner similar to thatdescribed for Example A(1). p-Tolyl isothiocyanate and2-bromo-2′,4′-dimethoxyacetophenone gave, after recrystallization fromMeOH/CHCl₃, 78 mg (24% yield) of a yellow solid, mp 215-216° C.

[0128]¹H NMR (DMSO-d₆): δ 10.51 (1H, s), 7.88 (2H, bs), 7.41 (2H, d,J=8.4 Hz), 7.16 (2H, d, J=8.4 Hz), 7.12 (2H, d, J=8.4 Hz), 6.58 (11H, d,J=2.2 Hz), 6.52 (1H, dd, J=8.4, 2.2 Hz), 3.78 (3H, s), 3.74 (3H, s),2.24 (3H, s).

[0129] IR(KBr): 3279, 2959, 1606, 1515, 1432, 1306, 1284, 1209, 1157,1124, 1032 cm⁻¹.

[0130] Anal. Calcd. for C₁₉H₁₉N₃O₃S: C, 61.77; H, 5.18; N, 11.37; S,8.68. Found: C, 61.69; H, 5.16; N, 11.33; S, 8.79.

Example A(14)[4-Amino-2-(p-tolylamino)-thiazol-5-yl]-(2,4-dimethylphenyl)-methanone

[0131]

[0132] The title compound was prepared in a manner similar to thatdescribed for Example A(1). p-Tolyl isothiocyanate and2-bromo-2′,4′-dimethylacetophenone gave, after recrystallization fromMeOH/CHCl₃, 65 mg (33% yield) of a yellow crystals, mp 220-221° C.

[0133]¹H NMR (DMSO-d₆): δ 10.58 (1H, s), 7.99 (2H, bs), 7.39 (2H, d,J=8.1 Hz), 7.17 (2H, d, J=7.8 Hz), 7.13 (2H, d, J=8.1 Hz), 7.04 (1H, s),7.00 (1H, d, J=7.8 Hz), 2.73 (3H, s), 2.24 (3H, s), 2.22 (3H, s).

[0134] IR(KBr): 3266, 2921, 1612, 1598, 1546, 1518, 1431 cm⁻¹.

[0135] Anal. Calcd. for C₁₉H₁₉N₃OS: C, 67.63; H, 5.68; N, 12.45; S,9.50. Found: C, 67.70; H, 5.73; N, 12.47; S, 9.62.

Example B [4-Amino-2-(p-tolylamino)-thiazole-5-carbonyl]-phenyl Benzoate

[0136]

[0137] An intermediate,S-(4-Benzoyloxyphenylacetyl)-N′-cyano-N″-p-tolyl-isothiourea, was firstprepared following a procedure in Gewald et al., J. Prakt. Chem, vol. 35(1967), pp. 97-104. Sodium (6.7 mg, 0.29 mmol) was carefully dissolvedin methanol (0.5 mL) and allowed to cool to ambient temperature. To theresultant solution was added p-tolyl isothiocyanate (43 mg, 0.29 mmol)and cyanamide (12 mg, 0.29 mmol). After 1 hour, 4-bromoacetylphenylbenzoate (92 mg, 0.29 mmol) was added, and the resultant mixture wasstirred overnight at ambient temperature. The mixture was then dilutedwith water (10 mL). The resulting tan solid was filtered off, rinsedwith water and a small amount of ether, dried under vacuum, andrecrystallized from ethanol/CHCl₃ to give an initial crop of 63 mg (51%yield) of S-(4-benzoyloxyphenylacetyl)-N′-cyano-N″-p-tolyl-isothiourea(as white needles):

[0138]¹H NMR (DMSO-d₆): δ 8.10-8.04 (2H, m), 7.72 (1H, ddd, J 7.5, 1.8,1.8 Hz), 7.67-7.54 (4H, m), 7.20 (2H, d, J=8.7 Hz), 7.03 (4H, dd,J=11.2, 8.7 Hz), 4.10 (1H, d, J=12.1 Hz), 3.77 (1H, d, J=12.1 Hz), 2.19(3H, s).

[0139] From the intermediate the title compound was prepared as follows.Crude S-(4-benzoyloxyphenylacetyl)-N′-cyano-N″-p-tolyl-isothiourea (0.29mmol) and triethylamine (101 μL, 0.73 mmol) in ethyl acetate (1 mL) washeated at reflux for 2 hours, then allowed to cool to ambienttemperature, and concentrated in vacuo to a crude solid, whichcrystallized from MeOH/CHCl₃ in successive crops to afford 67 mg (54%yield) of yellow needles, mp 245-247° C.

[0140]¹H NMR (DMSO-d₆): δ 10.71 (1H, s), 8.34-8.11 (4H, m), 7.80-7.72(3H, m), 7.66-7.57 (2H, m), 7.46 (2H, d, J=8.1 Hz), 7.38 (2H, d, J=8.4,Hz), 7.16 (2H, d, J=8.4 Hz), 2.26 (3H, s).

[0141] IR (KBr): 3451, 3332, 3026, 1732, 1597, 1522, 1444, 1264, 1206,1164, 1062, 708 cm⁻¹.

[0142] HRFABMS: Calcd. for C₂₄H₁₉N₃O₃SCs (M+Cs⁺): 562.0201. Found:562.0184.

[0143] Anal. Calcd. for C₂₄H₁₉N₃O₃S: C, 67.12; H, 4.46; N, 9.78; S,7.47. Found: C, 66.90; H, 4.43; N, 9.70; S, 7.50.

Example C(1) 4-[4-Amino-5-(4-methoxy-benzoyl)-thiazol-2-ylamino]-benzoicAcid Methyl Ester

[0144]

[0145] To a mixture of 4-methoxycarbonylphenyl isothiocyanate (82 mg,0.5 mmol) and cyanamide (23 mg, 0.55 mmol) in acetonitrile (5 mL), asolution of potassium tert-butoxide (61 mg, 0.55 mmol) in tert-butanol(5 mL) was added. After 30 minutes at ambient temperature,2-bromo-4′-methoxy-acetophenone (115 mg, 0.5 mmol) was added. After 2hours at ambient temperature, the reaction mixture was diluted withwater (50 mL). The product was collected by filtration, rinsed withwater and ethyl ether, and dried under vacuum to furnish a yellow solid,172 mg (90% yield).

[0146]¹H NMR (DMSO-d₆): δ 8.00 (2H, d, J=8.2 Hz), 7.84 (2H, d, J=8.2Hz), 7.72 (2H, d, J=8.2 Hz), 7.10 (2H, d, J=8.2 Hz), 3.90 (6H, s).

[0147] FABMS (MH⁺): 384.

[0148] Anal. Calcd. for C₁₉H₁₇N₃O₄S.0.5H₂O: C, 57.36; H, 4.71; N, 10.56;S, 8.06. Found: C, 56.97; H, 4.74; N, 10.51; S, 8.07.

Example C(2)[4-Amino-2-(4-benzyloxy-phenylamino)-thiazol-5-yl]-(4-methoxy-phenyl)-methanone

[0149]

[0150] The title compound was prepared in a manner like that describedfor Example C(1). 4-Benzyloxy-phenyl isothiocyanate and2-bromo-4′-methoxy-acetophenone gave a yellow-brown solid in 85% yield,mp 222-224° C.

[0151]¹H NMR (DMSO-d₆): δ 7.70 (2H, d, J=8.2 Hz), 7.58-7.34 (7H, m),7.06 (4H, dd, J=7.5, 1.2 Hz), 5.18 (2H, s), 3.94 (3H, s).

[0152] FABMS (MH⁺): 432.

[0153] Anal. Calcd. for C₂₄H₂₁N₃O₃S: C, 66.80; H, 4.91; N, 9.74; S,7.43. Found: C, 66.86; H, 4.91; N, 9.76; S, 7.53.

Example C(3)[4-Amino-2-(4-dimethylamino-phenylamino)-thiazol-5-yl]-(4-methoxy-phenyl)-methanone

[0154]

[0155] The title compound was prepared similarly as described forExample C(1) from 4-dimethylamino-phenyl isothiocyanate and2-bromo-4′-methoxy-acetophenone to give the product as a yellow solid in85% yield, mp 178-180° C.

[0156]¹H NMR (DMSO-d₆): δ 7.70 (2H, d, J=8.2 Hz), 7.34 (2H, d, J=8.2Hz), 7.00 (2H, d, J=8.6 Hz), 6.80 (2H, d, J=8.6), 3.94 (3H, s), 2.94(6H, s).

[0157] Anal. Calcd. for C₁₉H₂₀N₄O₂S: C, 61.94; H, 5.47; N, 15.21; S,8.70. Found: C, 62.22; H, 5.48; N, 15.03; S, 8.58.

Example C(4)[4-Amino-2-(4-dimethylamino-phenylamino)-thiazol-5-yl]-(2-nitro-phenyl)-methanone

[0158]

[0159] The title compound was prepared in a manner analogous to thatdescribed for Example C(1). 4-Dimethylamino-phenyl isothiocyanate and2-bromo-2′-nitro-acetophenone furnished a yellow solid in 90% yield,mp>195° C. (decomp.).

[0160]¹H NMR (DMSO-d₆): δ 8.06 (2H, bs), 7.76 (2H, m), 7.27 (2H, bs),6.74 (2H, d, J=9.0 Hz), 3.38 (6H, s).

[0161] FABMS (MH⁺): 384.

[0162] Anal. Calcd. for C₁₈H₁₇N₅O₃S.0.5 CH₃CN: C, 56.49; H, 4.62; N,19.07; S, 7.94. Found: C, 56.71; H, 4.64; N, 19.09; S, 7.88.

Example C(5)(4-Amino-2-phenethylamino-thiazol-5-yl)-(2-nitro-phenyl)-methanone

[0163]

[0164] The title compound was prepared essentially as described forExample C(1). Phenethyl isothiocyanate and 2-bromo-2′-nitro-acetophenoneprovided an amorphous yellow solid in 90% yield, mp 75.0-81.5° C.(decomp.).

[0165]¹H NMR (DMSO-d₆): δ 8.67 (1H, bs), 8.00 (1H, d, J=8.1 Hz), 7.80(2H, bs), 7.75 (1H, t, J=7.5 Hz), 7.65 (1H, t, J=7.5 Hz), 7.58 (1H, d,J=6.5 Hz), 7.04-7.32 (5H, m), 3.50 (2H, bs), 2.82 (2H, t, J=7.2 Hz).

[0166] FABMS (MH⁺): 369.

[0167] Anal. Calcd. for C₁₈H₁₆N₄O₃S.0.1H₂O.0.1 C₆H₁₄: C, 58.97; H, 4.68;N, 14.79; S, 8.46. Found: C, 58.97; H, 4.78; N, 14.54; S, 8.37.

Example C(6) Methyl2(S)-[4-Amino-5-(4-nitro-benzoyl)-thiazol-2-ylamino]-butyrate

[0168]

[0169] The title compound was prepared in a manner similar to thatdescribed for Example C(1). Methyl 2(S)-isothiocyanato-butyrate and2-bromo-4′-nitro-acetophenone afforded an amorphous red-brown solid in89% yield.

[0170]¹H NMR (CDCl₃): δ 8.28 (2H, d, J=8.2 Hz), 7.86 (2H, J=8.2 Hz),3.94 (3H, s), 4.32 (1H, bs), 2.12 (1H, m), 1.88 (1H, m), 0.96 (3H, t,J=6.4 Hz).

[0171] FABMS (MH⁺): 365.

Example C(7)[4-Amino-2-((4-dimethylaminophenyl)amino)-thiazol-5-yl]-(3-methyl-benzo[b]thiophen-2-yl)-methanone

[0172]

[0173] The title compound was prepared essentially as described forExample C(1). 4-Dimethylaminophenyl isothiocyanate and2-(2-bromoacetyl)-3-methyl-benzo[b]thiophene gave, afterrecrystallization from ethanol, 210 mg (92% yield) of yellow powder, mp123-126° C.

[0174]¹H NMR (DMSO-d₆): δ 10.50 (1H, s), 8.20 (2H, bs), 7.96 (1H, ddd,J=5.0, 5.0, 1.9 Hz), 7.82 (1H, ddd, J=4.1, 4.1, 1.7 Hz), 7.44 (2H, ddd,J=9.0, 4.5, 4.5 Hz), 7.26 (2H, d, J=8.5 Hz), 6.69 (2H, d, J=9.0 Hz),2.84 (6H, s).

[0175] FABMS (MH⁺): 409.

[0176] Anal. Calcd. for C₂₁H₂₀N₄OS₂.0.5H₂O.0.5 MeOH: C, 59.56; H, 5.35;N, 12.92; S, 14.79. Found: C, 59.87; H, 5.39; N, 12.86; S, 14.69.

Example C(8)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(3-methyl-benzo[b]thiophen-2-yl)-methanone

[0177]

[0178] The title compound was prepared in a manner like that describedfor Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al.,Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822) and2-(2-bromoacetyl)-3-methyl-benzo[b]thiophene provided 160 mg (53% yield)of yellow powder, mp 235-240° C.

[0179]¹H NMR(DMSO-d₆): δ 12.50 (1H, s), 10.9 (1H, s), 8.28 (2H, bs),8.19 (1H, s), 8.20-7.93 (1H, m), 7.93-8.00 (2H, m), 7.56 (1H, d, J=8.7Hz), 7.50-7.40 (2H, m), 7.25 (1H, d, J=8.7 Hz), 2.49 (3H, s).

[0180] FABMS (MH⁺): 406.

[0181] Anal. Calcd. for C₂₀H₁₅N₅OS₂.0.5H₂O: C, 57.95; H, 3.89; N, 16.90;S, 15.47. Found: C, 57.98; H, 3.88; N, 17.06; S, 15.55.

Example C(9)[4-Amino-2-(5-chloro-3-methyl-benzo[b]thiophen-2-ylamino)-thiazol-5-yl]-(4-dimethylamino-phenyl)-methanone

[0182]

[0183] The title compound was prepared essentially as described forExample C(1). 4-Dimethylaminophenyl isothiocyanate and2-(2-bromoacetyl)-5-chloro-3-methyl-benzo[b]thiophene provided 54% yieldof yellow powder, mp 265-268° C.

[0184]¹H NMR (DMSO-d₆): δ 10.60 (1H, s), 8.04 (2H, bs), 8.00 (1H, d,J=8.7 Hz), 7.88 (1H, d, J=1.8 Hz), 7.46 (1H, dd, J=8.7, 1.8 Hz),7.34-7.20 (2H, m), 6.68 (2H, d, J=9.0 Hz), 2.85 (6H, s), 2.43 (3H, s).

[0185] FABMS (MH⁺): 443/445.

[0186] Anal. Calcd. for C₂₁H₁₉N₄OS₂Cl: C, 56.94; H, 4.32; N, 12.65; S,14.48; Cl, 8.00. Found: C, 56.82; H, 4.39; N, 12.42; S, 14.42; Cl, 8.17.

Example C(10)[4-Amino-2-(1H-benzoimidazol-6-yl-amino)-thiazol-5-yl]-(5-chloro-3-methyl-benzo[b]thiophen-2-yl)-methanone

[0187]

[0188] The title compound was prepared in a similar manner as thatdescribed for Example C(1). 6-Isothiocyanato-1H-benzoimidazole (seeBoev, et al., Pharm. Chem. J. (Engl. Transl), vol. 24 (1990), pp.818-822) and 2-(2-bromoacetyl)-5-chloro-3-methyl-benzo[b]thiopheneprovided 59% yield of yellow powder, mp 275-280° C.

[0189]¹H NMR (DMSO-d₆): δ 12.44 (1H, s), 10.86 (1H, s), 8.30 (2H, bs),8.18 (1H, s), 8.02 (1H, d, J=8.4 Hz), 7.90 (1H, d, J=2.0 Hz), 7.86 (1H,bs), 7.55 (1H, d, J=8.4 Hz), 7.45 (1H, dd, J=8.7, 2.0 Hz), 7.25 (1H, d,J=8.7 Hz), 2.46 (3H, s).

[0190] ESIMS (MH⁺): 440/442.

[0191] Anal. Calcd. for C₂₀H₁₄N₅OS₂Cl.1.0H₂O: C, 52.45; H, 3.52; N,15.29; S, 14.00; Cl, 7.74. Found: C, 52.61; H, 3.60; N, 15.15; S, 14.12;Cl, 7.81.

Example C(11)[4-Amino-2-(benzo[1,3]dioxol-5-yl-amino)-thiazol-5-yl]-(2-nitro-phenyl)-methanone

[0192]

[0193] The title compound was prepared analogously to Example C(1).3,4-Methylenedioxyphenyl isothiocyanate and2-bromo-2′-nitro-acetophenone provided a yellow solid in 73% yield, mp200.0-202.5° C.

[0194]¹H NMR (DMSO-d₆): δ 10.69 (1H, s), 8.04 (2H, bs), 8.03 (1H, d,J=7.8 Hz), 7.78 (1H, dd, J=7.8, 6.5 Hz), 7.67 (1H, dd, J=7.2, 6.5 Hz),7.63 (1H, d, J=7.2 Hz), 7.28 (1H, s), 6.89 (1H, d, J=8.9 Hz), 6.85 (1H,d, J=8.9 Hz), 6.00 (2H, s).

[0195] FABMS (MH⁺): 385.

[0196] Anal. Calcd. for C₁₇H₁₂N₄O₅S: C, 53.12; H, 3.15; N, 14.58; S,8.34. Found: C, 53.02; H, 3.20; N, 14.39; S, 8.27.

Example C(12)[4-Amino-2-(4-methoxy-phenylamino)-thiazol-5-yl]-(2-iodo-phenyl)-methanone

[0197]

[0198] 2-Bromo-2′-iodoacetophenone, which has the structural formula

[0199] was first prepared as follows. According to a procedure of Kinget al., J. Org. Chem, vol. 29 (1964), pp. 3459-3461, to a solution of2′-iodoacetophenone (3.54 g, 14.4 mmol) in EtOAc was added copper(II)bromide (6.34 g, 28.8 mmol), and the resulting mixture was heated atreflux for 90 minutes. The mixture was allowed to cool, and the solidwas filtered off and rinsed with EtOAc. The filtrate was concentrated,providing 4.60 g (98% yield) of 2-bromo-2′-iodoacetophenone as a yellowoil, which matched previously reported material (Lutz et al., J. Org.Chem., vol. 12 (1947), p. 617).

[0200] The title compound was next prepared essentially as described forExample C(1). 4-Methoxyphenyl isothiocyanate and2-bromo-2′-iodoacetophenone provided a yellow solid in 71% yield, mp187-190° C.

[0201]¹H NMR (DMSO-d₆): δ 10.56 (1H, s), 8.03 (2H, bs), 7.85 (1H, d,J=7.5 Hz), 7.32-7.48 (3H, m), 7.29 (1H, dd, J=7.5, 1.6 Hz), 7.12 (1H,td, J=7.5, 1.6 Hz), 6.90 (2H, d, J=9.0 Hz), 3.51 (3H, s).

[0202] FABMS (MH⁺): 452.

[0203] Anal. Calcd. for C₁₇H₁₄N₃O₂SI.0.05 C₆H₆.0.2EtOH: C, 45.78; H,3.36; N, 9.05; S, 6.90; I, 27.33. Found: C, 46.06; H, 3.54; N, 9.09; S,7.04; I, 27.62.

Example C(13)[4-Amino-2-(4-nitro-phenylamino)-thiazol-5-yl]-(2-nitro-phenyl)-methanone

[0204]

[0205] The title compound was prepared in a manner similar to thatdescribed for Example C(1). 4-Nitrophenyl isothiocyanate and2-bromo-2′-nitroacetophenone provided a yellow solid in 45% yield, mp266.0-268.2° C.

[0206]¹H NMR (DMSO-d₆): δ 10.80 (1H, s), 8.23 (2H, d, J=9.4 Hz), 8.15(2H, bs), 8.08 (1H, d, J=8.1 Hz), 7.84 (2H, d, J=9.4 Hz), 7.83 (1H, t,J=7.5 Hz), 7.75-7.66 (2H, m).

[0207] FABMS (MH⁺): 386.

[0208] Anal. Calcd. for C₁₆H₁₁N₅O₅S: C, 49.87; H, 2.88; N, 18.17; S,8.32. Found: C, 49.83; H, 2.90; N, 18.10; S, 8.27.

Example C(14)(4-Amino-2-cyclohexylamino-thiazol-5-yl)-(2-nitro-phenyl)-methanone

[0209]

[0210] The title compound was prepared analogously to Example C(1).Cyclohexyl isothiocyanate and 2-bromo-2′-nitroacetophenone provided ayellow solid in 45% yield, mp 116-118° C.

[0211]¹H NMR (DMSO-d₆): δ 8.62 (1H, bs), 8.00 (1H, d, J=8.1 Hz), 7.97(2H, bs), 7.75 (1H, dd, J=8.1, 7.5 Hz), 7.64 (1H, dd, J=8.1, 7.5 Hz),7.59 (11H, d, J=7.5 Hz), 3.62 (1H, bs), 1.94-1.78 (2H, m), 1.73-1.60(2H, m), 1.58-1.46 (1H, m), 1.32-1.02 (5H, m).

[0212] FABMS (MH⁺): 347.

[0213] Anal. Calcd. for C₁₆H₁₈N₄O₃S.0.7H₂O: C, 53.53; H, 5.45; N, 15.61;S, 8.93. Found: C, 53.79; H, 5.24; N, 15.44; S, 8.93.

Example C(15)[4-Amino-2-(4-isopropyl-phenylamino)-thiazol-5-yl]-(2-nitro-phenyl)-methanone

[0214]

[0215] The title compound was prepared essentially as described forExample C(1). Isopropyl isothiocyanate and 2-bromo-2′-nitroacetophenoneprovided a yellow solid in 58% yield, mp 202.5-205.0° C.

[0216]¹H NMR (DMSO-d₆): δ 10.74 (1H, s), 8.05 (2H, bs), 8.03 (1H, d,J=7.5 Hz), 7.78 (1H, dt, J=7.5, 1.3 Hz), 7.71-7.60 (2H, m), 7.41 (2H, d,J=8.3 Hz), 7.20 (2H, d, J=8.3 Hz), 2.83 (1H, heptet, J=6.9 Hz), 1.16(6H, d, J=6.9 Hz).

[0217] FABMS (MH⁺): 383.

[0218] Anal. Calcd. for C₁₉H₁₈N₄O₃S: C, 59.67; H, 4.74; N, 14.65; S,8.38. Found: C, 59.62; H, 4.77; N, 14.56; S, 8.43.

Example C(16){4-Amino-2-[2-(4-chloro-phenyl)-ethylamino]-thiazol-5-yl}-(2-nitro-phenyl)-methanone

[0219]

[0220] The title compound was prepared in a manner similar to thatdescribed for Example C(1). 4-Chlorophenethyl isothiocyanate and2-bromo-2′-nitroacetophenone provided a yellow solid in 61% yield, mp117-120° C.

[0221]¹H NMR (DMSO-d₆): δ 8.74 (1H, bs), 8.00 (1H, d, J=8.1 Hz), 7.95(2H, bs), 7.75 (1H, td, J=7.5, 1.2 Hz), 7.64 (2H, td, J=8.1, 1.6 Hz),7.57 (1H, dd, J=7.5, 1.2 Hz), 7.33 (2H, d, J=8.4 Hz), 7.23 (2H, d, J=8.4Hz), 3.60-3.35 (2H, m), 2.81 (2H, t, J=6.8 Hz).

[0222] FABMS (MH⁺): 403.

[0223] Anal. Calcd. for C₁₈H₁₅N₄O₃SCl.0.5 EtOH: C, 53.58; H, 4.26; N,13.16; S, 7.53; Cl, 8.32. Found: C, 53.63; H, 4.33; N, 13.22; S, 7.47;Cl, 8.45.

Example C(17)[4-Amino-2-(4-diethylamino-phenylamino)-thiazol-5-yl]-(2-nitro-phenyl)-methanone

[0224]

[0225] The title compound was prepared in a manner like that describedfor Example C(1). 4-Diethylaminophenyl isothiocyanate and2-bromo-2′-nitroacetophenone provided a yellow solid in 63% yield, mp202.5-205.0° C.

[0226]¹H NMR (DMSO-d₆): δ 10.45 (1H, s), 8.01 (1H, d, J=8.1 Hz), 7.97(2H, bs), 7.75 (1H, dd, J=8.1, 7.8 Hz), 7.64 (11H, dd, J=8.1, 7.8 Hz),7.59 (1H, d, J=7.8 Hz), 7.18 (2H, d, J=9.0 Hz), 6.61 (2H, d, J=9.0 Hz),3.28 (4H, q, J=7.2 Hz), 1.05 (6H, t, J=7.2 Hz).

[0227] FABMS (MH⁺): 412.

[0228] Anal. Calcd. for C₂₀H₂₁N₅O₃S: C, 58.38; H, 5.14; N, 17.02; S,7.79. Found: C, 58.28; H, 5.20; N, 16.77; S, 7.94.

Example C(18)[4-Amino-2-(4-diethylamino-phenylamino)-thiazol-5-yl]-(4-nitro-phenyl)-methanone

[0229]

[0230] The title compound was prepared in a manner analogous to thatdescribed for Example C(1). 4-Diethylaminophenyl isothiocyanate and2-bromo-4′-nitroacetophenone provided a yellow solid in 63% yield, mp220-221° C.

[0231]¹H NMR (DMSO-d₆): δ 10.51 (1H, s), 8.42 (2H, bs), 8.26 (2H, d,J=12.0 Hz), 7.84 (2H, d, J=12.0 Hz), 7.22 (2H, d, J=9.0 Hz), 6.63 (2H,d, J=9.0 Hz), 3.26 (4H, q, J=6.8 Hz), 1.05 (6H, t, J=6.8 Hz).

[0232] FABMS (MH⁺): 412.

[0233] Anal. Calcd. for C₂₀H₂₁N₅O₃S: C, 58.38; H, 5.14; N, 17.02; S,7.79. Found: C, 58.23; H, 5.16; N, 16.94; S, 7.86.

Example C(19)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(3-methyl-thiophen-2-yl)-methanone

[0234]

[0235] The title compound was prepared essentially in the mannerdescribed for Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boevet al., Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822)and 2-(2-bromoacetyl)-3-methyl-thiophene (U.S. Pat. No. 5,189,049; anacetyl brominated with copper(II) bromide according to a procedure fromKing, et al., J. Org. Chem., Vol. 29 (1964), pp. 3459-3461;representative procedure in Example C(19)) provided 67% yield of yellowpowder, mp 285-287° C.

[0236]¹H NMR (DMSO-d₆): δ 12.60 (1H, bs), 10.78 (1H, s), 8.23 (1H, s),8.17 (2H, bs), 7.93 (1H, s), 7.56 (1H, d, J=8.7 Hz), 7.55 (1H, d, J=5.0Hz), 7.27 (1H, dd, J=8.7, 1.9 Hz), 6.60 (1H, d, J=5.0 Hz), 2.36 (3H, s).

[0237] FABMS (MH⁺): 356.

[0238] Anal. Calcd. for C₁₆H₁₃N₅OS₂.0.6H₂O: C, 52.47; H, 3.91; N, 19.12;S, 17.51. Found: C, 52.50; H, 3.90; N, 19.10; S, 17.71.

Example C(20)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(2,4-dimethyl-phenyl)-methanone

[0239]

[0240] The title compound was prepared in a manner similar to thatdescribed for Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boevet al., Pharm. Chem. J. (Engl. Transl.), vol. 24 (1990), pp. 818-822)and 2-bromo-2′,4′-dimethylacetophenone provided 77% yield of yellowpowder, mp 290-292° C.

[0241]¹H NMR (DMSO-d₆): δ 12.43 (1H, bs), 10.65 (1H, s), 8.18 (1H, s),8.00 (2H, bs), 7.80 (1H, s), 7.54 (1H, d, J=8.7 Hz), 7.20 (1H, d, J=8.7Hz), 7.16 (1H, d, J=7.5 Hz), 7.03 (1H, s), 6.99 (1H, d, J=7.5 Hz), 2.26(3H, s), 2.22 (3H, s).

[0242] FABMS (MH⁺): 364.

[0243] Anal. Calcd. for C₁₉H₁₇N₅OS: C, 62.79; H, 4.71; N, 19.27; S,8.82. Found: C, 62.50; H, 4.78; N, 19.22; S, 8.72.

Example C(21)[4-Amino-2-(pyridin-3-ylamino)-thiazol-5-yl]-(2,4-dimethyl-phenyl)-methanone

[0244]

[0245] The title compound was prepared in a manner similar to thatdescribed for Example C(1). 3-Pyridyl-isothiocyanate and2-bromo-2′,4′-dimethylacetophenone provided 63% yield of yellow powder,mp 200-202° C.

[0246]¹H NMR (DMSO-d₆): δ 10.82 (1H, s), 8.76 (1H, d, J=2.5 Hz), 8.25(1H, d, J=4.1 Hz), 8.06 (1H, d, J=8.4 Hz), 8.04 (2H, bs), 7.36 (1H, dd,J=8.4, 4.1 Hz), 7.21 (1H, d, J=7.5 Hz), 7.06 (1H, s), 7.02 (1H, d, J=7.5Hz), 2.28 (3H, s), 2.23 (3H, s).

[0247] FABMS (MH⁺): 325.

[0248] Anal. Calcd. for C₁₇H₁₆N₄OS: C, 62.94; H, 4.97; N, 17.27; S,9.88. Found: C, 62.86; H, 5.03; N, 17.17; S, 9.95.

Example C(22)3-[4-Amino-5-(2-cyano-benzoyl)-thiazol-2-ylamino]-benzonitrile

[0249]

[0250] The title compound was prepared essentially as described forExample C(1). 3-Cyanophenyl isothiocyanate and2-bromo-2′-nitro-acetophenone furnished an orange solid in 94% yield, mp235-236° C.

[0251]¹H NMR (DMSO-d₆): δ 8.26 (1H, bs), 8.06 (1H, d, J=8.0 Hz), 7.8(1H, t, J=7.0 Hz), 7.74-7.64 (3H, m), 7.58-7.48 (2H, m).

[0252] IR (KBr): 3460, 3307, 3271, 3083, 2214, 1625, 1601, 1525 cm⁻¹.

[0253] Anal. Calcd. For C₁₇H₁₁N₅O₃S: C, 55.80; H, 3.03; N, 19.17; S,8.78. Found: C, 55.70; H, 3.05; N, 19.01; S, 8.73.

Example C(23)[4-Amino-2-(3-methoxy-propylamino)-thiazol-5-yl]-(2-nitro-phenyl)-methanone

[0254]

[0255] The title compound was prepared analogously to Example C(1).3-Methoxypropyl isothiocyanate and 2-bromo-2′-nitro-acetophenonefurnished a yellow solid in 90% yield, mp 170-172° C.

[0256]¹H NMR (DMSO-d₆): δ 8.02-7.92 (2H, m), 7.4 (1H, t, J=7.0 Hz),7.68-7.56 (2H, m), 3.38-3.22 (7H, m), 1.78-1.66 (2H, m).

[0257] Anal. Calcd. for C₁₄H₁₆N₄O₄S: C, 49.99; H, 4.79; N, 16.66; S,9.53. Found: C, 50.04; H, 4.81; N, 16.69; S, 9.61.

Example C(24)1-{4-[4-Amino-5-(2-nitro-benzoyl)-thiazol-2-ylamino]-phenyl}-ethanone

[0258]

[0259] The title compound was prepared in a manner similar to ExampleC(1). 4-Acetylphenyl isothiocyanate and 2-bromo-2′-nitro-acetophenonefurnished a yellow solid in 87% yield, mp 264-265° C.

[0260]¹H NMR (DMSO-d₆): δ 8.06 (1H, d, J=8.0 Hz), 7.92 (2H, d, J=9.0Hz), 7.84-7.78 (1H, m), 7.73-7.64 (4H, m), 2.42 (3H, s).

[0261] IR (KBr): 3389, 3248, 1690, 1655, 1537, 1472, 1420, 1273 cm⁻¹.

[0262] Anal. Calcd. for C₁₈H₁₄N₄O₄S: C, 56.54; H, 3.69; N, 14.65; S,8.39. Found: C, 56.39; H, 3.73; N, 14.44; S, 8.31.

Example C(25) {4-Amino-2-[4-(2-chloro-5-trifluoromethyl-pyridin-2-ylsulfanyl)-phenylamino]-thiazol-5-yl}-(2-nitro-phenyl)-methanone

[0263]

[0264] The title compound was prepared in a manner similar to thatdescribed for Example C(1).2-[4-(2-Chloro-5-trifluoromethyl-pyridine-2-yl-sulfanyl)-phenyl]isothiocyanate and 2-bromo-2′-nitro-acetophenone furnished an orangesolid in 52% yield, mp 150-152° C.

[0265]¹H NMR (DMSO-d₆): δ 8.65 (1H, bs), 8.38 (1H, bs), 8.06 (2H, d,J=8.0 Hz), 7.80 (1H, t, J=7.0 Hz), 7.74-7.64 (4H, m), 7.54 (2H, d, J=8.0Hz).

[0266] IR (KBr): 3272, 3048, 1596, 1531, 1431, 1320 cm⁻¹.

[0267] Anal. Calcd. for C₂₂H₁₃ClF₃N₅O₃S₂: C, 47.87; H, 2.37; N, 12.69;S, 11.62; Cl, 6.42. Found: C, 47.79; H. 2.44; N, 12.54; S, 11.70; Cl,6.52.

Example C(26) Methyl3-[4-Amino-5-(2-methoxy-benzoyl)-thiazol-2-ylamino]-benzoate

[0268]

[0269] The title compound was prepared essentially as described forExample C(1). 3-Methoxycarbonylphenyl isothiocyanate and2-bromo-2′-methoxy-acetophenone gave an ivory solid in 59% yield, mp214-215° C.

[0270]¹H NMR (DMSO-d₆): δ 10.81 (1H, s), 8.12-7.90 (4H, m), 7.62 (1H,ddd, J=7.8, 1.2, 1.2 Hz), 7.49 (1H, t, J=7.9 Hz), 7.39 (1H, ddd, J=8.7,8.7, 1.7 Hz), 7.25 (1H, dd, J=7.5, 1.9 Hz), 7.09 (1H, d, J=8.4 Hz), 6.98(1H, ddd, J=7.5, 7.5, 0.6 Hz), 3.85 (3H, s), 3.87 (3H, s).

[0271] FABMS (MH⁺): 327.

[0272] IR (KBr): 3473, 3333, 3261, 3092, 1718, 1602, 1527, 1417, 1294cm⁻¹.

[0273] Anal. Calcd. for C₁₉H₁₇N₃O₄S: C, 59.52; H, 4.47; N, 10.96; S,8.36. Found: C, 59.41; H, 4.46; N, 10.93; S, 8.38.

Example C(27){4-Amino-2-[2-(4-chloro-phenyl)-ethylamino]-thiazol-5-yl}-(2-methoxy-phenyl)-methanone

[0274]

[0275] The title compound was prepared in a manner like that describedfor Example C(1). The product from 4-chlorophenethyl isothiocyanate and2-bromo-2′-methoxy-acetophenone was extracted with 10% i-PrOH/CHCl₃. Theresultant solid was washed with Et₂O to give an ivory solid in 49%yield, mp 150-151° C.

[0276]¹H NMR (DMSO-d₆): δ 8.53 (2H, bs), 7.87 (1H, bs), 7.39-7.28 (3H,m), 7.23 (2H, d, J=8.4 Hz), 7.17 (1H, dd, J=7.5, 1.6 Hz), 7.03 (1H, d,J=8.4 Hz), 6.93 (1H, t, J=7.5 Hz), 3.88 (3H, s), 3.40 (2H, bs), 2.81(2H, t, J=7.0 Hz).

[0277] FABMS (MH+): 388.

[0278] IR (KBr): 3354, 3214, 3166, 3103, 1600, 1578, 1544, 1525, 1462,1363 cm⁻¹.

[0279] Anal. Calcd. for C₁₉H₁₈ClN₃O₂S: C, 58.83; H, 4.68; N, 10.83; S,8.27. Found: C, 58.70; H, 4.62; N, 10.75; S, 8.25.

Example C(28)[4-Amino-2-(pyridin-3-ylamino)-thiazol-5-yl]-(2,4-dichloro-phenyl)-methanone

[0280]

[0281] The title compound was prepared in a manner similar to thatdescribed for Example C(1). 3-Pyridyl-isothiocyanate and2,2′,4′-trichloroacetophenone gave a yellow solid in 39% yield, mp209-2100° C.

[0282]¹H NMR (DMSO-d₆): δ 10.95 (11H, s), 8.77 (1H, d, J=2.5 Hz), 8.28(1H, dd, J=4.7, 1.6 Hz), 8.16 (2H, bs), 8.06 (1H, bd, J=9.6 Hz), 7.70(1H, d, J=1.6 Hz), 7.48 (2H, dd, J 11.5, 8.1 Hz), 7.37 (1H, dd, J=8.4,4.7 Hz).

[0283] FABMS (MH+): 365.

[0284] IR (KBr): 3378, 3272, 3175, 3072, 1608, 1586, 1561, 1525, 1424cm⁻¹.

[0285] Anal. Calcd. for C₁₅H₁₀Cl₂N₄OS.0.9H₂O: C, 47.23; H, 3.12; N,14.69; S, 8.41. Found: C, 47.03; H, 3.09; N, 14.52; S, 8.42.

Example C(29)[4-Amino-2-(pyridin-3-ylamino)-thiazol-5-yl]-(2-methoxy-phenyl)-methanone

[0286]

[0287] The title compound was prepared in a manner analogous to ExampleC(1). 3-Pyridyl-isothiocyanate and 2-bromo-2′-methoxy-acetophenone gavean off-white/ivory solid in 67% yield, mp 245-246° C.

[0288]¹H NMR ([DMSO-d₆): δ 10.80 (1H, s), 8.77 (1H, d, J=2.8 Hz), 8.25(1H, dd, J=4.7, 1.2 Hz), 8.07 (1H, ddd, J 8.4, 2.8, 1.6 Hz), 8.00 (2H,bs), 7.44-7.33 (211, m), 7.24 (11H, dd, J=7.5, 1.6 Hz), 7.09 (1H, d,J=8.1 Hz), 6.98 (1H, t, J=7.5 Hz), 3.76 (3H, s).

[0289] FABMS (MH+): 327.

[0290] IR (KBr): 3424, 3310, 2971, 1632, 1603, 1526, 1459, 1405 cm⁻¹.

[0291] Anal. Calcd. for C₁₆H₁₄N₄O₂S: C, 58.88; H, 4.32; N, 17.17; S,9.82. Found: C, 58.84; H, 4.33; N, 17.07; S, 9.90.

Example C(30)[4-Amino-2-(pyridin-3-ylamino)-thiazol-5-yl]-naphthalen-2-yl-methanone

[0292]

[0293] The title compound was prepared essentially as described forExample C(1). 3-Pyridyl-isothiocyanate and 2-bromo-2′-acetonaphthonegave, after recrystallization from EtOH, a yellow solid in 12% yield, mp242-243° C. (decomp.).

[0294]¹H NMR (DMSO-d₆): δ 10.97 (1H, s), 8.82 (1H, d, J=2.5 Hz),8.36-8.18 (3H, m), 8.13 (1H, ddd, J=8.4, 4.0, 1.6 Hz), 8.08-7.93 (2H,m), 7.77 (1H, dd, J=8.4, 1.6 Hz), 7.60 (2H, dddd, J=14.3, 10.6, 7.9, 2.2Hz), 7.39 (1H, dd, J=8.4, 5.0 Hz).

[0295] FABMS (MH⁺): 347.

[0296] IR (KBr): 3462, 3316, 3261, 3071, 1623, 1584, 1531, 1421 cm⁻¹.

[0297] Anal. Calcd. for C₁₉H₁₄N₄OS: C, 65.88; H. 4.07; N, 16.17; S,9.26. Found: C, 65.80; H, 4.09; N, 16.09; S, 9.34.

Example C(31)[4-Amino-2-(2-methoxy-benzylamino)-thiazol-5-yl]-(5-chloro-benzofuran-2-yl)-methanone

[0298]

[0299] The title compound was prepared in a manner similar to thatdescribed for Example C(1). 2-Methoxybenzyl isothiocyanate and2-bromoacetyl-5-chlorobenzofuran provided 62% yield of yellow powder, mp241-242° C.

[0300]¹H NMR (DMSO-d₆): δ 9.17 (1H, bs), 8.78 (1H, bs), 8.21 (1H, bs),7.83 (1H, d, J=2.2 Hz), 7.66 (1H, d, J=9.0 Hz), 7.44 (1H, dd, J=9.0, 2.2Hz), 7.39 (1H, s), 7.28 (1H, d, J=8.1 Hz), 7.25 (1H, dd, J=7.5, 7.2 Hz),7.01 (1H, d, J=8.1 Hz), 6.92 (1H, dd, J=7.5, 7.2 Hz), 4.51 (2H, bs),3.82 (3H, s).

[0301] FABMS (MH⁺): 414/416.

[0302] Anal. Calcd. for C₂₀H₁₆N₃O₃ClS: C, 58.04; H, 3.90; N, 10.15; S,7.75; Cl, 8.57. Found: C, 57.97; H, 3.85; N, 10.11; S, 7.85; Cl, 8.63.

Example C(32)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(2-methoxy-phenyl)-methanone

[0303]

[0304] The title compound was prepared in a manner analogous to thatdescribed for Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boevet al., Pharm. Chem. J. (Engl. Transl.), vol. 24 (1990), pp. 818-822)and 2-bromo-2′-methoxyacetophenone provided 72% yield of amorphousyellow powder, mp 180-185° C. (decomp.).

[0305]¹H NMR (DMSO-d₆): δ 12.40 (1H, bs), 10.61 (1H, bs), 8.16 (1H, s),7.94 (2H, bs), 7.83 (1H, bs), 7.53 (1H, d, J=8.4 Hz), 7.36 (1H, ddd,J=8.4, 7.6, 1.6 Hz), 7.24-7.16 (2H, m), 7.05 (1H, d, J=8.1 Hz), 6.95(1H, dd, J=7.6, 7.2 Hz), 3.74 (3H, s).

[0306] FABMS (MH⁺): 366.

[0307] Anal. Calcd. for C₁₈H₁₅N₅O₂S.0.5H₂O: C, 57.74; H, 4.31; N, 18.71;S, 8.56. Found: C, 57.78; H, 4.29; N, 18.64; S, 8.53.

Example C(33)4-[4-Amino-5-(2,4-dimethoxy-benzoyl)-thiazol-2-ylamino]-benzenesulfonamide

[0308]

[0309] The title compound was prepared essentially as described forExample C(1). 4-Isothiocyanato-benzenesulfonamide and2-bromo-2′,4′-dimethoxyacetophenone provided 75% yield of yellow powder,mp 249-250° C.

[0310]¹H NMR (DMSO-d₆): δ 10.93 (1H, bs), 7.93 (2H, bs), 7.75 (4H, bs),7.25 (2H, bs), 7.21 (1H, d, J=8.1 Hz), 6.61 (1H, d, J=1.9 Hz), 6.55 (1H,dd, J=8.1, 1.9 Hz), 3.79 (3H, s), 3.76 (3H, s).

[0311] FABMS (MH⁺): 435.

[0312] Anal. Calcd. for C₁₈H₁₈N₄O₅S₂: C, 49.76; H, 4.18; N, 12.89; S,14.76. Found: C, 49.66; H, 4.15; N, 12.77; S, 14.86.

Example C(34) Ethyl4-[4-Amino-2-(4-sulfamoyl-phenylamino)-thiazole-5-carbonyl]-benzoate

[0313]

[0314] The title compound was prepared substantially as described forExample C(1). 4-Isothiocyanato-benzenesulfonamide and ethyl4-bromoacetyl-benzoate provided 95% yield of yellow powder, mp 225-227°C.

[0315]¹H NMR (DMSO-d₆): δ 11.16 (1H, s), 8.32 (2H, bs), 8.04 (2H, d,J=8.4 Hz), 7.80 (2H, d, J=8.4 Hz), 7.78 (4H, bs), 7.26 (2H, bs), 4.33(2H, q, J=7.2 Hz), 1.33 (3H, t, J=7.2 Hz).

[0316] FABMS (MH⁺): 447.

[0317] Anal. Calcd. for C₁₉H₁₈N₄O₅S₂.0.4H₂O: C, 50.30; H, 4.18; N,12.38; S, 14.13. Found: C, 50.11; H, 3.97; N, 12.26; S, 14.14.

Example C(35)4-[4-Amino-5-(2,4-dimethyl-benzoyl)-thiazol-2-ylamino]-benzenesulfonamide

[0318]

[0319] The title compound was prepared essentially as described forExample C(1). 4-Isothiocyanatobenzenesulfonamide and2-bromo-2′,4′-dimethylacetophenone furnished a yellow solid in 75%yield, mp 242-244° C.

[0320]¹H NMR (DMSO-d₆): δ 10.97 (1H, bs), 8.00 (2H, bs), 7.76 (2H, d,J=9.7 Hz), 7.72 (2H, d, J=9.7 Hz), 7.24 (2H, bs), 7.22 (1H, d, J=7.5Hz), 7.07 (1H, s), 7.03 (1H, d, J=7.5 Hz), 2.29 (3H, s), 2.23 (3H, s).

[0321] FABMS (MH⁺): 403.

[0322] Anal. Calcd. for C₁₈H₁₈N₄O₃S₂: C, 53.71; H, 4.51; N, 13.92; S,15.93. Found: C, 53.47; H, 4.54; N, 13.69; S, 15.83.

Example C(36) {4-Amino-2-[4-(2-chloro-5-trifluoromethyl-pyridine-2-ylsulfanyl)-phenylamino]-thiazol-5-yl}-(2,6-dichloro-4-trifluoromethyl-phenyl)-methanone

[0323]

[0324] The title compound was prepared essentially as described forExample C(1).2-[4-(2-Chloro-5-trifluoromethyl-pyridin-2-yl-sulfanyl)-phenyl]isothiocyanate and 2-bromo-2′,6′dichloro-4′-trifluoromethyl-acetophenonefurnished an orange solid in 52% yield, mp 130-132° C.

[0325]¹H NMR (DMSO-d₆): δ 8.65 (1H, bs), 8.38 (1H, bs), 8.06 (2H, d,J=8.0 Hz), 7.80 (1H, t, J=7.0 Hz), 7.74-7.64 (4H, m), 7.54 (2H, d, J=8.0Hz).

[0326] IR (KBr): 3272, 3048, 1596, 1531, 1431, 1320 cm⁻¹.

[0327] Anal. Calcd. for C₂₂H₁₃ClF₃N₅O₃S₂: C, 47.87; H, 2.37; N, 12.69;S, 11.62; Cl, 6.42. Found: C, 47.79; H, 2.44; N, 12.54; S, 11.70; Cl,6.52.

Example C(37)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(2,6-dichloro-4-trifluoromethyl-phenyl)-methanone

[0328]

[0329] The title compound was prepared in a manner analogous to thatdescribed for Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boevet al,. Pharm. Chem. J. (Engl. Transl.), vol. 24 (1990), pp. 818-822)and 2-bromo-2′,6′-dichloro-4′-trifluoromethyl-acetophenone gave a yellowsolid in 56% yield that decomposed at a temperature above 180° C.

[0330]¹H NMR (DMSO-d₆): δ 12.45 (1H, bd, J=16.0 Hz), 11.10-10.80 (1H,m), 8.20 (1H, s), 8.00 (2H, s), 7.70-7.45 (2H, m), 7.20 (1H, d, J=8.0Hz).

[0331] IR (KBr): 3191, 2974, 1619, 1559, 1467, 1309 cm⁻¹.

[0332] FABMS (MH+): 472.

[0333] Anal. Calcd. for C₁₈H₁₀C₁₂F₃N₅OS.0.6 HOAc.0.1 CH₂Cl₂. H₂O: C,45.58; H, 2.95; N, 12.18; S, 5.69; Cl, 13.92. Found: C, 45.70; H, 3.05;N, 12.45; Cl, 13.87.

Example C(38)4-[4-Amino-5-(2,6-dichloro-4-trifluoromethyl-benzoyl)-thiazol-2-ylamino]-benzenesulfonamide

[0334]

[0335] The title compound was prepared essentially as described forExample C(1). 4-Isothiocyanatobenzenesulfonamide and2-bromo-2′,6′-dichloro-4′-(trifluoromethyl) acetophenone furnished,after recrystallization from EtOH/H₂O and drying via benzene azeotrope,a yellow solid in 46% yield, mp 294-296° C.

[0336]¹H NMR (DMSO-d₆): δ 8.10 (1H, s), 8.05 (2H, s) 7.77 (4H, dd,J=9.0, 14.0 Hz).

[0337] HRFABMS: Calcd. for C₇H₁₂Cl₂F₃N₄O₃S₂(MH⁺): 510.9680. Found:510.9697.

[0338] Anal. Calcd. for C₁₇H₁₁Cl₂F₃N₄O₃S₂.0.1H₂O. C₆H₆: C, 40.28; H,2.51; N, 10.30; S, 11.97; Cl, 13.51. Found: C, 40.58; H, 2.28; N, 10.75;S, 12.31; Cl, 13.61.

Example C(39) Phenyl4-[4-Amino-2-(4-sulfamoyl-phenylamino)-thiazole-5-carbonyl]-benzoate

[0339]

[0340] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Isothiocyanato-benzenesulfonamide and4-(bromoacetyl)-phenyl benzoate provided a yellow solid in 77% yield,mp>300° C.

[0341]¹H NMR (DMSO-d₆): δ 11.13 (1H, s), 8.26 (2H, bs), 8.15 (2H, dd,J=7.2, 1.6 Hz), 7.83-7.73 (7H, m), 7.66-7.59 (2H, m), 7.41 (2H, d, J=6.9Hz), 7.27 (2H, s).

[0342] HRFABMS (MH⁺): Calcd.: 495.0797. Found: 495.0812.

[0343] Anal. Calcd. for C₂₃H₁₈N₄O₅S₂.0.2H₂O: C, 55.45; H, 3.72; N,11.25; S, 12.87. Found: C, 55.34; H, 3.592; N, 11.01; S, 12.88.

Example C(40)[4-Amino-2-(4-methoxy-phenylamino)-thiazol-5-yl]-(4-methyl-1H-imidazol-5-yl)-methanone

[0344]

[0345] 5-Bromoacetyl-4-methyl-1H-imidazole, which has the structuralformula

[0346] was first prepared as follows. Bromine (0.40 mL, 7.77 mmol) wasadded dropwise to a solution of 5-acetyl-4-methyl-1H-imidazole (964 mg,7.77 mmol; LaMattina et al, J. Org. Chem., vol. 48 (1983), pp. 897-898)in HOAc (20 mL). After two days, the HOAc was removed in vacuo and theresidue partitioned with CH₂Cl₂ and sat. aq. NaHCO₃. The organic layerwas washed with brine, dried over Na₂SO₄, and evaporated to provide alight brown solid, 625 mg (40% yield), which was used without furtherpurification.

[0347]¹H NMR (DMSO-d₆): δ 12.65 (1H, bs), 7.67 (1H, s),4.62 (2H, s),2.44 (3H, s).

[0348] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Methoxy-phenyl isothiocyanate and5-bromoacetyl-4-methyl-1H-imidazole provided a yellow powder in 57%yield, mp 248-50° C.

[0349]¹H NMR (DMSO-d₆): δ 12.28 (1H, bs), 10.21 (1H, s), 8.00 (2H, bs),7.56 (1H, s), 7.49 (2H, d, J=9.0 Hz), 6.94 (2H, d, J=9.0 Hz), 3.75 (3H,s), 2.50 (3H, s).

[0350] HRFABMS (M+Na⁺): Calcd.: 352.0844. Found: 352.0840.

[0351] Anal. Calcd. for C₁₅H₁₅N₅O₂S.0.5H₂O: C, 53.24; H, 4.77; N, 20.70;S, 9.48. Found: C, 53.43; H, 4.78; N, 20.54; S, 9.38.

Example C(41)[4-Amino-2-(4-imidazol-1-yl-phenylamino)-thiazol-5-yl]-(2,4-dimethyl-phenyl)-methanone

[0352]

[0353] 1-(4-Isothiocyanato-phenyl)-1H-imidazole, which has thestructural formula

[0354] was first prepared as follows. To a solution of1-(4-amino-phenyl)-1H-imidazole (1.00 g, 6.30 mmol; Venuti et al, J.Med. Chem., vol. 31 (1988), pp. 2136-2145) in acetone (10 mL) at 0° C.was simultaneously added a solution of thiophosgene (580 μL, 7.6 mmol)in acetone (15 mL) and a solution of 25% aq. Na₂CO₃ (15 mL). The mixturewas stirred at 0° C. for 0.5 hour and allowed to warm to roomtemperature over 1.5 hour. The acetone was removed under reducedpressure and the residue diluted with H₂O. The cream-colored precipitatewas filtered off, washed with H₂O, and dried under high vacuum to give1.20 g (95% crude yield) of a light tan solid, which was used withoutfurther purification.

[0355]¹H NMR (DMSO-d₆): δ 8.33 (1H, s), 7.81 (1H, s), 7.76 (2H, d, J=8.8Hz), 7.61 (2H, d, J=8.8 Hz), 7.12 (1H, s).

[0356] The title compound was prepared in a manner like that describedfor Example C(1). 1-(4-Isothiocyanato-phenyl)-1H-imidazole and2-bromo-2′,4′-dimethyl-acetophenone provided a yellow solid in 14%yield, mp 180.0-180.5° C.

[0357]¹H NMR (DMSO-d₆): δ 10.80 (1H, s), 8.10 (1H, s), 8.02 (1H, bs),7.68 (2H, d, J=7.5 Hz), 7.58 (2H, d, J=9.0 Hz), 7.20 (1H, d, J=7.8 Hz),7.10-7.00 (2H, m), 2.28 (3H, s), 2.24 (3H, s).

[0358] IR (KBr): 3393, 3119, 2925, 1612, 1566, 1524, 1425 cm⁻¹.

[0359] FABMS (MH⁺): 390.

[0360] Anal. Calcd. for C₂₁H₁₉N₅OS.0.2H₂O: C, 64.17; H. 4.97; N, 17.82;S, 8.16. Found: C, 64.14; H, 4.98; N, 17.68; S, 8.21.

Example C(42)[4-Amino-2-(4-imidazol-1-yl-phenylamino)-thiazol-5-yl]-(3-methyl-thiophen-2-yl)-methanone

[0361]

[0362] The title compound was prepared in a manner like that describedfor Example C(1). 1-(4-Isothiocyanato-phenyl)-1H-imidazole (from ExampleC(41)) and 2-bromoacetyl-3-methyl-thiophene (from Example C(19))provided a yellow solid in 83% yield, mp>300° C.

[0363]¹H NMR (DMSO-d₆): δ 10.98 (1H, s), 8.25 (1H, s), 8.18 (1H, bs),7.77 (1H, s), 7.72 (2H, J=6.5 Hz), 7.65 (1H, s), 7.62 (2H, J=4.7), 7.10(1H, s), 6.98 (1H, d, J=5.0 Hz), 2.28 (3H, s).

[0364] IR (KBr): 3402, 3278, 3103, 2982, 1609, 1523, 1422, 1306 cm⁻¹.

[0365] Anal. Calcd. for C₁₇H₁₆N₅OS₂: C, 56.67; H, 3.96; N, 18.36; S,16.81. Found: C, 56.38; H, 4.06; N, 18.13; S, 16.67.

Example C(43)[4-Amino-2-(1H-benzimidazol-5-ylamino)-thiazol-5-yl]-(1-methyl-1H-pyrrol-2-yl)-methanone

[0366]

[0367] The title compound was prepared in a manner analogous to thatused in Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev etal., Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822) and2-chloroacetyl-N-methyl-pyrrole (Croce et al., Synthesis (1990), pp.212-213) provided a yellow solid in 42% yield, mp 284-285° C.

[0368]¹H NMR (DMSO-d₆): δ 12.43 (1H, bs), 10.65 (1H, bs), 8.18 (1H, s),7.94 (3H, bs), 7.55 (1H, d, J=8.7 Hz), 7.27 (11H, dd, J=8.7, 1.9 Hz),6.92 (1H, m), 6.62 (11H, dd, J=3.7, 2.1 Hz), 6.04 (1H, dd, J=4.1, 2.1Hz), 3.80 (3H, s).

[0369] HRFABMS (MH⁺): Calcd.: 339.1028. Found: 339.1024.

[0370] Anal. Calcd. for C₁₆H₁₄N₆OS.0.3H₂O: C, 55.90; H, 4.28; N, 24.45;S, 9.33. Found: C, 56.08; H, 4.28; N, 24.46; S, 9.33.

Example C(44)1-{4-[4-Amino-5-(3-methyl-thiophene-2-carbonyl)-thiazol-2-ylamino]-phenyl}-ethanone

[0371]

[0372] The title compound was prepared essentially as described forExample C(1). 4-Acetylphenyl isothiocyanate and2-bromoacetyl-3-methyl-thiophene (from Example C(19)) gave a yellowsolid in 89% yield, mp 171-2° C.

[0373]¹H NMR (DMSO-d₆): δ 11.14 (1H, s), 8.22 (2H, bs), 7.95 (2H, d,J=9.0 Hz), 7.76 (2H, d, J=9.0 Hz), 7.62 (1H, d, J=5.0 Hz), 7.00 (1H, d,J=5.0 Hz), 2.53 (3H, s), 2.39 (3H, s).

[0374] IR (KBr): 3618, 3354, 3254, 3178, 3072, 1651, 1599, 1524, 1403,1355, 1318, 1275, 1170 cm⁻¹.

[0375] FABMS (MH⁺): 357.

[0376] Anal. Calcd for C₁₇H₁₅N₃O₂S₂.0.5H₂O: C, 55.72; H, 4.40; N, 11.47;S, 17.50. Found: C, 55.92; H, 4.44; N, 11.51; S, 17.44.

Example C(45)trans-3RS-Amino-4RS-{4-[4-amino-5-(3-methyl-thiophene-2-carbonyl)-thiazol-2-ylamino]-benzoyl}-dihydro-furan-2-one

[0377]

[0378] The title compound was prepared essentially as described forExample C(1). The product from 4-isothio-cyanato-benzoyl-DL-homoserinelactone and 2-bromoacetyl-3-methyl-thiophene (from Example C(19)) wasextracted into 10% i-PrOH/CHCl₃. Flash column chromatography with2-3-4-5-6% MeOH/CH₂Cl₂ stepwise gradient gave a yellow solid in 43%yield, mp 162-3° C.

[0379]¹H NMR (DMSO-d₆): δ 11.05 (1H, s), 8.88 (2H, d, J=8.1 Hz), 8.32(2H, bs), 7.85 (2H, d, J=9.0 Hz), 7.74 (2H, d, J=9.0 Hz), 7.61 (1H, d,J=5.0 Hz), 6.99 (1H, d, J=5.0 Hz), 4.73 (1H, q, J=9.3 Hz), 4.40 (1H,ddd, J=10.8, 8.7, 2.0 Hz), 4.26 (1H, ddd, J=10.2, 8.7, 6.7 Hz).

[0380] IR (KBr): 3413, 3284, 3084, 1773, 1637, 1608, 1524, 1413, 1313,1254, 1181 cm⁻¹.

[0381] FABMS (MH⁺): 443.

[0382] Anal. Calcd for C₂₀H₁₈N₄O₄S₂.0.4H₂O: C, 53.41; H, 4.21; N, 12.46;S, 14.26. Found: C, 53.56; H, 4.28; N, 12.30; S, 14.43.

Example C(46) Ethyl3RS-[4-Amino-5-(3-methyl-thiophene-2-carbonyl)-thiazol-2-ylamino]-butyrate

[0383]

[0384] The title compound was prepared essentially as described forExample C(1). The product from ethyl dl-3-isothiocyanato-butyrate and2-bromoacetyl-3-methyl-thiophene (from Example C(19)) was extracted with10% i-PrOH/CHCl₃. Flash column chromatography with 3% MeOH/CH₂Cl₂ gave ayellow solid in 45% yield, mp 129-30° C.

[0385]¹H NMR (DMSO-d₆): δ 8.61 (1H, d, J=7.8 Hz), 8.08 (2H, bs), 7.53(1H, d, J=5.0 Hz), 6.94 (1H, d, J=5.0 Hz), 4.05 (2H, q, J=7.2 Hz), 2.33(3H, s), 1.22-1.12 (6H, m).

[0386] IR(KBr): 3307, 3213, 3160, 2976, 1737, 1618, 1586, 1526, 1423,1349, 1215, 1183, 1091 cm⁻¹.

[0387] FABMS (MH⁺): 353.

[0388] Anal. Calcd for C₁₅H₁₉N₃O₃S₂: C, 50.97; H, 5.42; N, 11.89; S,18.14. Found: C, 50.81; H, 5.39; N, 11.72; S, 17.97.

Example C(47)4-[4-Amino-5-(4-methyl-thiazole-5-carbonyl)-thiazol-2-ylamino]-benzenesulfonamide

[0389]

[0390] 5-Bromoacetyl-4-methyl-thiazole, which has the structural formula

[0391] was prepared as described in Sych et al., J. Gen. Chem. USSR,vol. 32 (1962), pp. 970-975. Bromine (0.75 mL, 7.77 mmol) was addeddropwise into the solution of 1-(4-methyl-thiazol-5-yl)-ethanone (2.05mg, 14.5 mmol; Ganapathi et al., Proc.-Indian Acad. Sci. Sect. A, vol.22 (1945), pp. 362-378) in HOAc (3 mL). The mixture was stirred at 85°C. for 1.5 hours and turned into yellow cake. HOAc (3 mL) was added, andafter 1.5 hours, allowed to cool. The HOAc was removed in vacuo and theresidue partitioned between CH₂Cl₂ and sat aq NaHCO₃. The organic layerwas washed with brine, dried over Na₂SO₄, and evaporated to give a blacksolid, 1.3 g (41% yield), which was used without further purification.

[0392]¹H NMR (CDCl₃): δ 8.85 (1H, s), 4.28 (2H, s), 2.81 (3H, s).

[0393] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Isothiocyanato-benzenesulfonamide and5-bromoacetyl-4-methyl-thiazole provided a brown solid in 31% yield, mp265-266° C.

[0394]¹H NMR (DMSO-d₆): δ 11.18 (1H, s), 9.08 (1H, s), 8.30 (2H, bs),7.78 (4H, bs), 7.72 (2H, bs), 2.55 (3H, s).

[0395] Anal. Calcd. for C₁₆H₁₃N₅O₃S₃: C, 42.52; H, 3.31; N, 17.11; S,24.32. Found: C, 42.28; H, 3.33; N, 17.15; S, 24.52.

Example C(48)4-[4-Amino-5-(3-methyl-thiophene-2-carbonyl)-thiazol-2-ylamino]-benzenesulfonamide

[0396]

[0397] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Isothiocyanato-benzenesulfonamide and2-bromoacetyl-3-methyl-thiophene (from Example C(19)) provided a yellowsolid in 69% yield, mp 284.5-286.0° C.

[0398]¹H NMR (DMSO-d₆): δ 11.11 (1H, s), 8.20 (2H, bs), 7.80 (2H, d,J=10.7 Hz), 7.76 (2H, d, J=10.7 Hz), 7.61 (1H, d, J=5.0 Hz), 7.26 (2H,s), 6.90 (1H, d, J=5.0 Hz), 2.38 (3H, s).

[0399] Anal. Calcd. for C₁₅H₁₄N₄O₃S₃: C, 45.67; H, 3.58; N, 14.20; S,24.39. Found: C, 45.52; H, 3.58; N, 14.04; S, 24.36.

Example C(49):4-[4-Amino-5-(3-methyl-benzo[b]thiophene-2-carbonyl)-thiazol-2-ylamino]-benzenesulfonamide

[0400]

[0401] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Isothiocyanato-benzenesulfonamide and2-(2-bromoacetyl)-3-methyl-benzo[b]thiophene provided a yellow powder in73% yield, mp 274.0-275.5° C.

[0402]¹H NMR (DMSO-d₆): δ 11.17 (1H, bs), 8.33 (2H, bs), 8.04-7.97 (1H,m), 7.90-7.84 (1H, m), 7.78 (4H, bs), 7.51-7.44 (2H, m), 7.27 (2H, s),2.52 (3H, s).

[0403] Anal. Calcd. for C₁₉H₁₆N₄O₃S₃: C, 51.33; H, 3.63; N, 12.60; S,21.64. Found: C, 51.19; H, 3.67; N, 12.31; S, 21.37.

Example C(50)4-[4-Amino-5-(2,5-dimethyl-thiophene-3-carbonyl)-thiazol-2-ylamino]-benzenesulfonamide

[0404]

[0405] 3-Bromoacetyl-2,5-dimethyl-thiophene, which has the structuralformula

[0406] was prepared in a manner analogous to 2-bromo-2′-iodoacetophenonefor Example C(12). 3-Acetyl-2,5-dimethylthiophene (6.83 g, 44.3 mmol)provided 10.1 g (98% yield) of yellow oil, which was used withoutfurther purification.

[0407]¹H NMR (CDCl₃): δ 7.22 (1H, s), 4.64 (2H, s), 2.58 (3H, s), 2.36(3H, s).

[0408] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Isothiocyanato-benzenesulfonamide and3-bromoacetyl-2,5-dimethyl-thiophene provided a yellow powder in 69%yield, mp 263-5° C.

[0409]¹H NMR (DMSO-d₆): δ 11.02 (1H, s), 8.05 (2H, bs), 7.76 (4H, s),7.25 (2H, s), 6.87 (1H, s), 2.43 (3H, s), 2.38 (3H, s).

[0410] Anal. Calcd. for C₁₆H₁₆N₄O₃S₃: C, 47.04; H, 3.95; N, 13.71; S,23.55. Found: C, 47.01; H, 3.92; N, 13.62; S, 23.47.

Example C(51)4-[4-Amino-5-(2-oxo-1,2,3,4-tetrahydro-quinoline-6-carbonyl)-thiazol-2-ylamino]-benzenesulfonamide

[0411]

[0412] The title compound was prepared essentially as described forExample C(1). 4-Isothiocyanato-benzenesulfonamide and6-(bromoacetyl)-2-oxo-1,2,3,4-tetrahydroquinoline gave a grey-yellowsolid in 48% yield, mp 300-305° C.(d).

[0413]¹H NMR (DMSO-d₆): δ 11.08 (1H, s), 10.32 (1H, s), 8.17 (2H, bs),7.82-7.70 (4H, m), 7.58-7.45 (3H, m), 7.27 (1H, s), 6.90 (1H, d, J=8.1Hz), 2.93 (4H, t, J=7.7 Hz).

[0414] IR (KBr): 3266, 3193, 3069, 1679, 1597, 1525, 1434, 1365, 1317,1153 cm⁻¹.

[0415] HRFABMS. Calcd. for C₁₉H₁₈N₅O₄S₂ (MH⁺): 444.0800. Found:444.0816.

[0416] Anal. Calcd for C₁₉H₁₇N₅O₄S₂.0.6 MeOH: C, 50.88; H, 4.23; N,15.13; S, 13.86. Found: C, 51.02; H, 4.00; N, 15.00; S, 13.60.

Example C(52)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(2,6-dichloro-phenyl)-methanone

[0417]

[0418] 2-Bromo-2′,6′-dichloro-acetophenone, which has the structuralformula

[0419] was prepared as follows. To 2′,6′-dichloroacetophenone (1.0 g,5.30 mmol) in HOAc (5 mL) was added dropwise bromine (272 μl, 5.30mmol). The mixture was heated at 90° C. for 1 hour, then diluted withice-water and partitioned between ether and sat aq NaHCO₃. The organiclayer was washed with brine, dried over MgSO₄, concentrated andazeotroped with heptane twice, to obtain 1.41 g (100% yield) of a lightyellow oil, which matched by ¹H NMR and IR previously described (seeMlotkowska et al., Pol. J. Chem., vol. 55 (1981), pp. 631-642) and wasused without further purification.

[0420]¹H NMR (CDCl₃): δ 7.39-7.33 (3H, m), 4.23 (2H, s).

[0421] The title compound was prepared in a manner analogous to thatused in Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev etal., Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822) and2-bromo-2′,6′-dichloro-acetophenone (from Example C(52)) provided ayellow solid in 47% yield, mp 203-208° C.

[0422]¹H NMR (DMSO-d₆): δ 12.47 (1H, d, J=17.7 Hz), 10.83 (1H, d, J=16.5Hz), 8.22-7.80 (3H, m), 8.18 (1H, s), 7.76-7.36 (5H, m), 7.19 (1H, d,J=8.4 Hz).

[0423] Anal. Calcd. for C₁₇H₁₂N₅OSCl₂: C, 50.51; H, 2.74; N, 17.32; S,7.93; Cl, 17.54. Found: C, 50.32; H, 2.78; N, 17.11; S. 7.91; Cl, 17.75.

Example C(53){4-Amino-2-[4-(1H-imidazol-2-yl)-phenylamino]-thiazol-5-yl}-(2,6-dichloro-4-trifluoromethyl-phenyl)-methanone

[0424]

[0425] 2-(4-Nitro-phenyl)-1H-imidazole, which has the structural formula

[0426] was first prepared as follows. To a solution of 2-phenylimidazole(5.00 g, 34.7 mmol) in conc. H₂SO₄ (20 mL) at 0° C. was added a solutionof conc. HNO₃ (2.2 mL, 35 mmol) in conc. H₂SO₄ (5 mL). The resultantbrown mixture was stirred at 0° C. for 2 hours and quenched withcrushed-ice. A pale white precipitate formed, which was filtered. Thefiltrate was brought to pH 9 with 2N NaOH. A yellow precipitate formed,which was filtered off, washed with H₂O, and recrystallized from boilingMeOH to give 3.0 g (46% yield) of a yellow solid. This crude product wasused without any further purification.

[0427]¹H NMR (MeOH-d₄): δ 8.34 (2H, d, J=9.0 Hz), 8.08 (2H, d, J=9.0Hz), 7.26 (2H, s).

[0428] 4-(1H-Imidazol-2-yl)-aniline, which has the structural formula

[0429] was next prepared as follows. To a suspension of2-(4-nitro-phenyl)-1H-imidazole (1.5 g, 7.93 mmol) in absolute ethanol(30 mL) was added 10% Pd—C (250 mg). The resultant mixture was stirredunder an atmosphere of H₂ for 5 hours. The mixture was filtered througha pad of Celite. The filtrate was concentrated under reduced pressure toafford 1.20 g (95% in crude yield) of a red gum, which was used withoutfurther purification.

[0430] 2-(4-Isothiocyanato-phenyl)-1H-imidazole, which has thestructural formula

[0431] was prepared in a manner analogous to1-(4-isothiocyanato-phenyl)-1H-imidazole for Example C(41).4-(1H-Imidazol-2-yl)-aniline gave a pale-brown solid, whichrecrystallized from CHCl₃ in 85% yield, and was used without any furtherpurification.

[0432]¹H NMR (MeOH-d₄): δ 7.88 (4H, bd, J=7.8 Hz), 7.58 (2H, s).

[0433] The title compound was prepared in a manner like that describedfor Example C(1). 2-(4-Isothiocyanato-phenyl)-1H-imidazole and2-bromo-2′,6′-dichloro-4′-trifluoromethyl-acetophenone gave, afterpurification via preparative thin layer chromatography with MeOH:CHCl₃(8:92) as eluant, a yellow solid in 21% yield, mp 195-197° C.

[0434]¹H NMR (DMSO-d₆): δ 11.0 (1H, s), 8.18 (1H, s), 8.02 (2H, s), 7.88(2H, d, J=8.7 Hz), 7.62 (2H, d, J=8.1 Hz), 7.12 (2H, bs).

[0435] IR (KBr): 3400, 2929, 1610, 1527, 1426, 1310 cm⁻¹.

[0436] HRFABMS: Calcd. for C₂₀H₁₃Cl₂F₃N₅OS (MH⁺): 498.0170. Found:498.0183.

[0437] Anal. Calcd. for C₂₀H₁₂Cl₂F₃N₅OS.H₂O: C, 46.52; H, 2.73; N,13.56; Cl, 13.73; S, 6.21. Found: C, 46.45; H, 2.78; N, 13.40; Cl,13.73, S, 6.11.

Example C(54)[4-Amino-2-(4-morpholin-4-yl-phenylamino)-thiazol-5-yl]-(2,4-dimethyl-phenyl)-methanone

[0438]

[0439] 4-(4-Isothiocyanato-phenyl)-morpholine, which has the structuralformula

[0440] was made as follows. To 4-morpholinoaniline (2.0 g, 11.2 mmol)and triethylamine (5.01 mL, 35.9 mmol) in THF (200 mL) at 0° C. wasadded dropwise thiophosgene (1.03 mL, 13.5 mmol). The mixture stirred atambient temperature overnight, and then was partitioned between etherand water. The ether layer was washed with water and brine, dried overMgSO₄, and concentrated to give 2.46 g (99%) of dark brown solid.

[0441]¹H NMR (CDCl₃): δ 7.15 (2H, d, J=9.3 Hz), 6.87 (2H, d, J=9.3 Hz),3.80 (4H, t, J=5.0 Hz), 3.19 (4H, t, J=5.0 Hz).

[0442] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-(4-Isothiocyanato-phenyl)-morpholine and2-bromo-2′,4-dimethylacetophenone provided a yellow solid in 28% yield,mp 253-254.5° C.

[0443]¹H NMR (DMSO-d₆): δ 10.44 (1H, s), 7.98 (2H, bs), 7.31 (2H, d,J=9.0 Hz), 7.14 (1H, d, J=7.8 Hz), 7.02 (1H, s), 6.99 (1H, d, J=7.8 Hz),6.90 (2H, d, J=9.0 Hz), 3.70 (4H, t, J=4.7 Hz), 3.04 (4H, t, J=4.7 Hz),2.26 (3H, s), 2.20 (3H, s).

[0444] Anal. Calcd. for C₂₂H₂₄N₄O₂S: C, 64.68; H, 5.92; N, 13.71; S,7.85. Found: C, 64.49; H, 5.97; N, 13.64; S, 7.93.

Example C(55)[4-Amino-2-(4-morpholin-4-yl-phenylamino)-thiazol-5-yl]-(2,6-dichloro-phenyl)-methanone

[0445]

[0446] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-(4-Isothiocyanato-phenyl)-morpholine (fromExample C(54)) and 2-bromo-2′,6′-dichloro-acetophenone (from ExampleC(52)) provided a yellow solid in 9% yield, mp 245-247° C.

[0447]¹H NMR (DMSO-d₆): δ 10.58 (1H, s), 8.02 (2H, bs), 7.52 (2H, d,J=7.3 Hz), 7.41 (1H, m), 7.30 (2H, d, J=9.0 Hz), 6.92 (2H, d, J=9.0 Hz),3.72 (4H, dd, J=5.0, 4.2 Hz), 3.06 (4H, dd, J 5.0, 4.2 Hz).

[0448] Anal. Calcd. for C₂₀H₁₈N₄O₂SCl: C, 53.46; H, 4.04; N, 12.47; S,7.14; Cl, 15.78. Found: C, 53.39; H, 4.04; N, 12.47; S, 7.21; Cl, 15.71.

Example C(56) Ethyl4-[4-Amino-5-(2,6-dichloro-benzoyl)-thiazol-2-ylamino]-benzoate

[0449]

[0450] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Ethoxycarbonylphenyl isothiocyanate and2-bromo-2′,6′-dichloro-acetophenone (from Example C(52)) provided anamorphous yellow solid in 48% yield.

[0451]¹H NMR (DMSO-d₆): δ 11.13 (1H, s), 8.15 (2H, bs), 7.92 (2H, d,J=8.7 Hz), 7.70 (2H, d, J=8.7 Hz), 7.58-7.40 (3H, m), 4.27 (2H, q, J=7.0Hz), 1.29 (3H, t, J=7.0 Hz).

[0452] Anal. Calcd. for C₁₉H₁₅N₃O₃SCl₂: C, 52.30; H, 3.47; N, 9.63; S,7.35; Cl, 16.25. Found: C, 52.20; H, 3.42; N, 9.63; S, 7.44; Cl, 16.26.

Example C(57)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(2,4,6-trimethyl-phenyl)-methanone

[0453]

[0454] 2-Bromo-2′,4′,6′-trimethyl-acetophenone, which has the structuralformula

[0455] was prepared in a manner analogous to2-bromo-2′-iodo-acetophenone, see Example C(12).2,4,6-trimethylacetophenone (1.50 g, 9.25 mmol) provided 2.26 g (100%)of clear oil, which was used without further purification.

[0456]¹H NMR (CDCl₃): δ 6.87 (2H, s), 4.27 (2H, s), 2.22 (9H, s).

[0457] The title compound was prepared in a manner analogous to thatused in Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev etal., Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822) and2-bromo-2′,4′,6′-trimethyl-acetophenone provided a yellow powder in 26%yield, that decomposed above 185° C.

[0458]¹H NMR (DMSO-d₆): δ 12.42 (1H, bs), 10.66 (1H, bs), 8.17 (1H, s),7.96 (2H, bs), 7.75 (1H, bs), 7.44 (1H, bs), 7.16 (1H, d, J=8.7 Hz),6.82 (2H, s), 2.21 (3H, s), 2.11 (6H, s).

[0459] HRFABMS (MH⁺): Calcd.: 378.1389. Found: 378.1381.

[0460] Anal. Calcd. for C₂₀H₁₉N₅OS.0.3H₂O: C, 62.74; H, 5.16; N, 18.29;S, 8.37. Found: C, 62.96; H, 5.14; N, 18.24; S, 8.35.

Example C(58)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(2,3,6-trimethyl-phenyl)-methanone

[0461]

[0462] 2-Bromo-2′,3′,6′-trimethyl-acetophenone, which has the structuralformula

[0463] was prepared in a manner analogous to2-bromo-2′-iodo-acetophenone for Example C(12).2′,3′,6′-trimethylacetophenone (1.50 g, 9.25 mmol) provided 2.10 g (93%)of clear oil, which was used without further purification.

[0464] The title compound was prepared in a manner analogous to thatused in Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev etal., Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822) and2-bromo-2′,3′,6′-trimethyl-acetophenone provided a yellow powder in 70%yield, that decomposed above 196° C.

[0465]¹H NMR (DMSO-d₆): δ 12.41 (1H, bs), 10.65 (1H, bs), 8.17 (1H, s),7.96 (2H, bs), 7.70 (1H, bs), 7.52 (1H, bs), 7.17 (1H, dd, J=8.4, 1.9Hz), 6.82 (2H, s), 2.11 (9H, s).

[0466] Anal. Calcd. for C₂₀H₁₉N₅OS: C, 63.64; H, 5.07; N, 18.55; S,8.50. Found: C, 63.40; H, 5.17; N, 18.37; S, 8.36.

Example C(59)[4-Amino-2-(4-sulfamoyl-phenylamino)-thiazol-5-yl]-(3,5-dimethyl-pyridin-4-yl)-methanone

[0467]

[0468] 4-(Bromoacetyl)-3,5-dimethylpyridine hydrobromide, which has thestructural formula

[0469] was first prepared as follows. 4-Acetyl-3,5-dimethylpyridine (500mg, 3.36 mmol; Kutney et al., Can. J. Chem., vol. 41 (1963), pp.695-702) was dissolved in 30% HBr in acetic acid (1 mL), heated to 70°C., and treated with a mixture of bromine (0.17 mL, 3.36 mmol) in 30%HBr in acetic acid (0.5 mL). After 2 hours, the mixture was allowed tocool to ambient temperature and ether (8 mL) was added. The resultantprecipitate was filtered off, rinsed with ether (2×), and dried toafford 1.03 g (100%) of a purple solid, mp 222-225° C., that was usedwithout further purification.

[0470] The title compound was prepared essentially as described forExample C(1). 4-Isothiocyanato-benzenesulfonamide and4-(bromoacetyl)-3,5-dimethylpyridine hydrobromide provided a tan solid,which was purified via column chromatography with 10% MeOH/CHCl₃ andcrystallized from MeOH to obtain 35 mg (51%) of amorphous yellow solid.

[0471]¹H NMR (DMSO-d₆): δ 11.09 (1H, s), 8.32 (2H, s), 8.18 (2H, bs),7.74 (4H, dd, J=11.5, 9.3 Hz), 7.27 (2H, s), 2.15 (6H, s).

[0472] IR (KBr): 3378, 3342, 3260, 3160, 1625, 1594, 1560, 1518, 1443,1342, 1160 cm⁻¹.

[0473] HRFABMS: Calcd. for C₁₇H₁₈N₅O₃S₂ (MH⁺): 404.0851. Found:404.0840.

[0474] Anal. Calcd for C₁₇H₁₇N₅O₃S₂.0.4H₂O.0.3 MeOH: C, 49.44; H, 4.56;N, 16.66; S, 15.26. Found: C, 49.13; H, 4.31; N, 16.61; S. 15.10.

Example C(60)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(2,6-dimethyl-phenyl)-methanone

[0475]

[0476] 2′,6′-Dimethylacetophenone, which has the structural formula

[0477] was prepared according to a procedure for o-nitro-acetophenone(Reynolds et al, Org. Syn. Coll., vol. IV (1963), pp. 708-710).2,6-Dimethylbenzoic acid (3.00 g, 20.0 mmol) provided 2.56 g (86% yield)of yellow oil, which was used without further purification.

[0478]¹H NMR (CDCl3): δ 7.16 (1H, t, J=7.2 Hz), 7.02 (2H, d, J=7.2 Hz),2.48 (3H, s), 2.25 (6H, s).

[0479] 2-Bromo-2′,6′-dimethyl-acetophenone, which has the structuralformula

[0480] was prepared in a manner analogous to2-bromo-2′-iodo-acetophenone, see Example C(12).2′,6′-dimethylacetophenone (1.50 g, 10.1 mmol) provided 2.04 g (89%yield) of clear oil, which was used without further purification.

[0481]¹H NMR (CDCl₃): δ 7.21 (1H, t, J=7.2 Hz), 7.05 (2H, t, J=7.2 Hz),4.29 (2H, s), 2.26 (6H, s).

[0482] The title compound was prepared in a manner analogous to thatused in Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev etal., Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822) and2-bromo-2′,6′-dimethyl-acetophenone provided a yellow solid in 71%yield, that decomposed above 185° C.

[0483]¹H NMR (DMSO-d₆): δ 12.41 (1H, bs), 10.67 (1H, bs), 8.17 (1H, s),7.99 (2H, s), 7.60 (1H, s), 7.52 (1H, s), 7.17 (1H, dd, J=8.7, 1.9 Hz),7.12 (1H, d, J=7.1 Hz), 7.02 (1H, d, J=7.5 Hz), 2.15 (6H, s).

[0484] HRFABMS (MH⁺): Calcd.: 364.1232. Found: 364.1227.

[0485] Anal. Calcd. for C₁₉H₁₇N₅OS.0.3CH₃OH: C, 62.14; H, 4.92; N,18.77; S, 8.60. Found: C, 62.43; H, 5.15; N, 18.91; S, 8.60.

Example C(61)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(2-methyl-6-nitro-phenyl)-methanone

[0486]

[0487] 2′-Methyl-6′-nitro-acetophenone, which has the structural formula

[0488] was prepared according to a procedure for o-nitro-acetophenone(see Reynolds et al, Org. Syn. Coll., vol. IV, (1963), pp. 708-710).2-Methyl-6-nitrobenzoic acid (15.0 g, 82.8 mmol) provided 14.7 g (99%yield) of yellow oil, which was used without further purification.

[0489]¹H NMR (CDCl₃): δ 8.04 (1H, d, J=8.4 Hz), 7.55 (1H, d, J=7.5 Hz),7.44 (1H, dd, J=8.4, 7.5 Hz), 2.56 (3H, s), 2.35 (3H, s).

[0490] 2-Bromo-2′-methyl-6′-nitro-acetophenone, which has the structuralformula

[0491] was prepared in a manner analogous to5-bromoacetyl-4-methyl-1H-imidazole for Example C(40). Crude2′-methyl-6′-nitro-acetophenone (1.56 g, 8.72 mL) furnished a whitesolid, 2.17 g (97% yield), that was used without further purification.

[0492]¹H NMR (CDCl₃): δ 8.11 (1H, d, J=7.8 Hz), 7.62 (1H, d, J=7.8 Hz),7.52 (1H, d, t=7.8 Hz), 4.33 (2H, s), 2.40 (3H, s).

[0493] The title compound was prepared in a manner analogous to thatused in Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev etal., Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822) and2-bromo-2′-methyl-6′-nitro-acetophenone provided a brown solid in 32%yield, mp 198-201° C.

[0494]¹H NMR (DMSO-d₆): δ 12.40 (1H, bs), 10.78 (1H, bs), 8.17 (1H, d,J=10.6 Hz), 8.00 (2H, bs), 7.92 (2H, d, J=8.4 Hz), 7.68 (1H, d, J=7.5Hz), 7.62-7.44 (2H, m), 7.19 (1H, d, J=7.5 Hz), 2.30 (3H, s).

[0495] HRFABMS (MH⁺): Calcd.: 395.0926. Found: 395.0920.

[0496] Anal. Calcd. for C₁₈H₁₄N₆O₃S.0.5H₂O: C, 53.59; H, 3.75; N, 20.83;S, 7.95. Found: C, 53.43; H, 3.67; N, 20.68; S, 7.81.

Example C(62)[4-Amino-2-(4-morpholin-4-yl-phenylamino)-thiazol-5-yl]-(2,6-dimethyl-phenyl)-methanone

[0497]

[0498] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-(4-Isothiocyanato-phenyl)-morpholine (fromExample C(54)) and 2-bromo-2′,6′-dimethyl-acetophenone (from ExampleC(60)) provided a brown solid in 23% yield, mp 221-223° C.

[0499]¹H NMR (DMSO-d₆): δ 10.42 (1H, s), 7.95 (2H, bs), 7.30 (2H, d,J=9.0 Hz), 7.18-7.10 (1H, m), 7.02 (2H, d, J=7.5 Hz), 6.91 (2H, d, J=9.0Hz), 3.72 (4H, t, J=4.8 Hz), 3.05 (4H, t, J=4.8 Hz), 2.16 (6H, s).

[0500] HRFABMS (M+): Calcd.: 408.1620. Found: 408.1607.

[0501] Anal. Calcd. for C₂₂H₂₄N₄O₂S.0.75H₂O: C, 62.61; H, 6.09; N,13.28; S, 7.60. Found: C, 62.64; H, 6.10; N, 13.05; S, 7.55.

Example C(63)[4-Amino-2-(1H-benzoimidazol-5-yl-amino)-thiazol-5-yl]-(3,5-dichloro-pyridin-4-yl)-methanone

[0502]

[0503] 4-Bromoacetyl-3,5-dichloropyridine, which has the structuralformula

[0504] was first prepared as follows. A mixture of3,5-dichloropyridine-4-carboxylic acid (4.00 g, 20.9 mmol; Cale et al.,J. Med. Chem., vol. 32 (1989), pp. 2178-2199), benzene (20 mL), DMF (0.4mL), and thionyl chloride (3.80 mL, 52.0 mmol) was heated at reflux for60 min, allowed to cool to ambient temperature, concentrated in vacuo,suspended in ether (20 mL), and cautiously treated with a solution oftrimethylsilyldiazomethane (25 mL of 2.0 M in hexanes). After 72 hours,48% HBr (18 mL) was carefully added dropwise over 20 min, initially withvigorous gas evolution. After 30 min, the mixture was made alkalinecarefully with NaHCO₃ and extracted with ether. The ethereal layers weredried over Na₂SO₄ and evaporated to give an orange oil, which waspurified via column chromatography with 50% CH₂Cl₂/hex eluant toseparate 2.50 g (51%) of 3,5-dichloropyridine-4-carbonyl chloride as ayellow oil, providing desired product, 2.00 g (36%) of pale yellowcrystals that darkened at ambient temperature, which was used withoutfurther purification.

[0505] NMR (CDCl₃): δ 8.58 (2H, s), 4.37 (2H, s).

[0506] Anal. Calcd for C₇H₄BrCl₂NO.0.02 C₆H₁₄: C, 31.60; H, 1.59; N,5.18. Found: C, 31.92; H, 1.59; N, 5.24.

[0507] The title compound was prepared essentially as described forExample C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al.,Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822) and4-(bromoacetyl)-3,5-dichloropyridine gave a product that was extractedinto 10% MeOH/CHCl₃ and column chromatography with same to furnish ayellow amorphous solid, 198 mg (55%). An analytical sample precipitatedfrom EtOH, mp 235-240° (d).

[0508]¹H NMR (CD₃OD): δ 8.60 (2H, s), 8.18 (1H, s), 7.98 (1H, bs), 7.58(1H, d, J=9.0 Hz), 7.30 (1H, dd, J=1.2, 8.7 Hz).

[0509] IR (KBr): 3183, 1608, 1544, 1461, 1427, 1355 cm⁻¹.

[0510] HRFABMS: Calcd. for C₁₆H₁₁Cl₂N₆OS (MH⁺): 405.0092. Found:405.0079.

[0511] Anal. Calcd for C₁₆H₁₀Cl₂N₆OS.1.1H₂O: C, 45.21; H, 2.89; N,19.77; Cl, 16.68; S, 7.54. Found: C, 45.49; H, 2.59; N, 19.64; Cl,16.62; S, 7.43.

Example C(64)2S-[4-Amino-2-(1H-benzoimidazol-5-yl-amino)-thiazole-5-carbonyl]-N-carbobenzyloxy-pyrrolidine

[0512]

[0513] 2S-Bromoacetyl-N-carbobenzyloxy-pyrrolidine, which has thestructural formula

[0514] was first prepared as follows. The acid chloride ofN-carbobenzyloxy-L-proline (1.20 g, 4.80 mmol) was made according toAoyama et al. Chem. Pharm. Bull., vol. 29 (1981), pp. 3249-3255, withoxalyl chloride and a catalytic amount of DMF. To a solution of thecrude acid chloride in THF (5 mL) and MeCN (5 mL) at 0° C. was carefullyadded dropwise a solution of trimethylsilyldiazomethane (5.0 mL of 2.0 Min hex), and initially vigorous gas evolution occurred. The resultantred suspension was allowed to warm and stirred at ambient temperatureovernight. The brown mixture was then cooled to 0° C., cautiouslytreated with a mixture of 47% HBr (4.1 mL) and ether (10 mL), andinitially vigorous gas evolution ensued. The mixture was allowed to warmto ambient temperature over 1 h, then made alkaline with satd aq NaHCO₃(20 mL), and extracted with EtOAc (2×20 mL). The combined organic layerswere dried over Na₂SO₄ and evaporated to give a brown oil, 1.57 g(100%), which was used without further purification.

[0515] NMR (CDCl₃): δ 7.44-7.24 (5H, m), 4.34 (1H, d, J=15.6 Hz), 4.27(1H, d, J=15.6 Hz).

[0516] The title compound was prepared essentially as described forExample C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al.,Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822) and2S-bromoacetyl-N-carbobenzyloxy-pyrrolidine provided a solid that wasprecipitated from iPrOH/hex twice to give a yellow amorphous solid, 154mg (54%), mp 150-165° (d).

[0517]¹H NMR (DMSO-d₆): δ 12.40 (1H, d, J=7.8 Hz), 10.68 (1H, d, J=19.3Hz), 8.20 (1H, d, J=10.6 Hz), 8.10-7.70 (2H, m), 7.52 (1H, dd, J=8.7,34.8 Hz), 7.45-7.05 (5H, m), 5.17-4.80 (2H, m), 4.32 (1H, d, J=4.9 Hz),4.30-4.18 (1H, bm), 2.33-1.70 (2H, bm).

[0518] IR (KBr): 3278, 1686, 1599, 1560, 1421, 1356, 1121 cm⁻¹.

[0519] HRFABMS: Calcd. for C₂₃H₂₃N₆O₃S (MH⁺): 463.1552. Found: 463.1538.

[0520] Anal. Calcd for C₂₃H₂₂N₆O₃S.0.1H₂O.0.7iPrOH: C, 59.53; H, 5.53;N, 16.60; S, 6.33. Found: C, 59.53; H, 5.53; N, 16.60; S, 6.22.

Example C(65)2S-[4-Amino-2-(4-sulfamoyl-phenylamino)-thiazole-5-carbonyl]-N-carbobenzyloxy-pyrrolidine

[0521]

[0522] The title compound was prepared essentially as described forExample C(1). 4-Isothiocyanato-benzenesulfonamide and2S-bromoacetyl-N-carbobenzyloxy-pyrrolidine (see Example C(64)) provideda solid that was purified via column chromatography with 5% MeOH/CHCl₃eluant to give a yellow amorphous solid, 140 mg (46%), mp 150-160° (d).

[0523]¹H NMR (DMSO-d₆): δ 11.05 (1H, d, J=10.0 Hz), 7.98 (2H, bd, J=17.1Hz), 7.79 (4H, dd, J=12.1, 9.7 Hz), 7.41-7.11 (5H, m), 5.15-4.89 (2H,m), 4.32-4.21 (1H, bm), 3.51-3.40 (2H, bm), 2.35-2.13 (1H, bm),1.93-1.75 (3H, bm).

[0524] IR(KBr): 3288, 1686, 1598, 1550, 1527, 1420, 1157 cm⁻¹.

[0525] HRFABMS: Calcd. for C₂₂H₂₃N₅O₅S₂Cs (M+Cs⁺): 634.0195. Found:634.0215.

[0526] Anal. Calcd for C₂₂H₂₃N₅O₅S₂.0.3H₂O.0.1CHCl₃: C, 51.15; H, 4.60;N, 13.50; S, 12.36. Found: C, 51.36; H, 4.63; N, 13.31; S, 12.47.

Example C(66) [4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(2-bromo-6-methyl-phenyl)-methanone

[0527]

[0528] 2′-Bromo-6′-methyl-acetophenone, which has the structural formula

[0529] was prepared in a manner analogous to o-nitro-acetophenone (seeReynolds et al., Org. Syn. Coll, vol. IV (1963), pp. 708-710). From2-methyl-6-bromobenzoic acid (3.10 g, 14.4 mmol) was provided 2.45 g(80%) of yellow oil, which matched previously described material by ¹HNMR (Swenton et al., J. Org. Chem., vol. 58 (1993), pp. 3308-3316) andwas used without further purification.

[0530] 2,2′-Dibromo-6′-methyl-acetophenone, which has the structuralformula

[0531] was prepared in a manner analogous to2-bromo-2′-iodo-acetophenone, see Example C(12). Crude2′-bromo-6′-methyl-acetophenone (1.00 g, 4.69 mmol) provided 1.48 g ofyellow oil, which was used without further purification.

[0532]¹H NMR (CDCl₃): δ 7.44-7.37 (1H, m), 7.21-7.17 (2H, m), 4.42 (2H,s), 2.31 (3H, s).

[0533] The title compound was prepared in a manner analogous to thatused in Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev etal., Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822) and2,2′-dibromo-6′-methyl-acetophenone provided a brown solid in 32% yield,mp 208-210° C.

[0534]¹H NMR (DMSO-d₆): δ 12.43 (1H, bs), 10.74 (1H, bs), 8.18 (1H, s),8.02 (2H, s), 7.75 (1H, bs), 7.44 (1H, bs), 7.44 (1H, d, J=7.5 Hz),7.28-7.14 (3H, m), 2.22 (3H, s).

[0535] ESIMS(MH⁺): 428/430.

[0536] Anal. Calcd. for C₁₈H₁₄N₅OSBr.1.0H₂O: C, 48.44; H, 3.61; N,15.69; S, 7.18; Br, 17.90. Found: C, 48.54; H, 3.69; N, 15.57; S, 7.11;Br, 17.88.

Example C(67)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(3-methyl-biphenyl-2-yl)-methanone

[0537]

[0538] 1-(3-Methyl-biphenyl-2-yl)-ethanone, which has the structuralformula

[0539] was prepared in the following manner. To2′-bromo-6′-methyl-acetophenone (from Example C(66); 760 mg, 3.58 mmol)and Pd(OAc)₂ (114 mg) in DMF (38 mL) at 0° C. under Ar° was added insuccession phenylboronic acid (495 mg) and 2M aq Na₂CO₃ (1.6 mL). Themixture was heated at 90° C. for 3 hours, then diluted with water (50mL), and extracted with ether (2×100 mL). The ethereal extracts wereconcentrated to a crude product, which was purified via columnchromatography with 2-5% ether/hexane stepwise gradient to obtain 670 mg(89% yield) of yellow oil, used without further purification.

[0540]¹H NMR (CDCl₃): δ 7.44-7.31 (5H, m), 7.25-7.19 (2H, m), 7.16-7.09(1H, m), 2.33 (3H, s), 1.93 (3H, s).

[0541] 2-Bromoacetyl-3-methyl-biphenyl, which has the structural formula

[0542] was prepared in a manner analogous to2-bromo-2′-iodo-acetophenone, see Example C(12). Crude1-(3-methyl-biphenyl-2-yl)-ethanone (295 mg, 1.40 mmol) provided 413 mgof yellow oil, which was used without further purification.

[0543]¹H NMR (CDCl3): δ 7.48-7.18 (8H, m), 4.42 (2H, s), 2.38 (3H, s).

[0544] The title compound was prepared in a manner analogous to thatused in Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev etal., Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822) and2-bromoacetyl-3-methyl-biphenyl provided a yellow solid in 49% yield, mp184-190° C.

[0545]¹H NMR (DMSO-d₆): δ 8.13 (1H, s), 7.87 (1H, s), 7.53 (1H, d, J=8.7Hz), 7.46-7.39 (2H, m), 7.38-7.15 (7H, m), 2.35 (3H, s). HRFABMS (M+):Calcd.: 426.1389. Found: 426.1374.

[0546] Anal. Calcd. for C₂₄H₁₉N₅OS.1.0H₂O.0.3CH₃CN: C, 64.82; H, 4.84;N, 16.29; S, 7.03. Found: C, 64.88; H, 4.69; N, 16.40; S, 7.28.

Example C(68)[4-Amino-2-(4-methoxy-benzylamino)-thiazol-5-yl]-(2,5-dimethylthiophen-3-yl)-methanone

[0547]

[0548] The title compound was prepared in a manner like that describedfor Example C(1). 3-Bromoacetyl-2,5-dimethyl-thiophene (from ExampleC(52)) and 1-(2-isothiocyanato-ethyl)-4-methoxy-benzene provided a whitesolid in 72% yield, mp 175° C.

[0549]¹H NMR (DMSO-d₆): δ 6.88 (2H, d, J=8.7 Hz), 6.74 (2H, d, J=8.7Hz), 6.41 (1H, s), 6.24 (1H, s), 4.88 (2H, s), 3.78 (3H, s), 2.40 (3H,s), 1.98 (3H, s).

[0550] IR (KBr): 3311, 2920, 1663, 1552, 1514, 1244 cm⁻¹.

[0551] FABMS (MH⁺): 380.

[0552] Anal. Calcd. for C₁₈H₁₉N₃O₂S₂: C, 57.88; H, 5.13; N, 11.25; S.17.17. Found: C, 57.97; H, 5.11; N, 11.33; S, 17.28.

Example C(69){4-Amino-2-[4-morpholin-4-yl-phenylamino]-thiazol-5-yl}-(3,5-dichloro-pyridin-4-yl)-methanone

[0553]

[0554] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-(4-Isothiocyanato-phenyl)-morpholine (fromExample C(54)) and 4-bromoacetyl-3,5-dichloro-pyridine (from ExampleC(63)) provided a yellow solid in 58% yield, mp 291.5-292.5° C.

[0555]¹H NMR (DMSO-d₆): δ 10.75 (1H, s), 8.71 (2H, s), 8.32 (1H, bs),8.01 (1H, bs), 7.30 (2H, bs), 6.92 (2H, d, J=9.0 Hz), 3.70 (4H, t, J=4.5Hz), 3.05 (4H, t, J=4.5 Hz).

[0556] FABMS (MH⁺): 450/452

[0557] Anal. Calcd. for C₁₉H₁₇N₅O₂SCl₂: C, 50.67; H, 3.80; N, 15.55; S,7.12, Cl, 15.74. Found: C, 50.55; H, 3.83; N, 15.29; S, 6.95, Cl, 15.47.

Example C(70){4-Amino-2-[4-(4-methyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(3,5-dichloro-pyridin-4-yl)-methanone

[0558]

[0559] 1-Methyl-4-(4-nitro-phenyl)-piperazine, which has the structuralformula

[0560] was first prepared as follows. A mixture of 1-methyl-piperazine(4.00 g, 39.9 mmol) and 1-chloro-4-nitro-benzene (3.14 g, 20.0 mmol) washeated to 80° C. for 24 hours, allowed to cool, and diluted with H₂O.The aqueous layer was extracted with MeOH:CH₂Cl₂ (20:80; 4×50 mL). Thecombined organic layers were dried over MgSO₄, filtered, concentratedunder reduced pressure, and recrystallized from ethanol to afford 3.2 g(75% yield) of a yellow solid, which matched previously reportedmaterial by ¹H NMR (de Silva et al., J. Chem. Soc. Perkin Trans. 2, vol.9 (1993), pp. 1611-1616) and was used without further purification.

[0561] 4-(4-Methyl-piperazin-1-yl)-aniline, which has the structuralformula

[0562] was next prepared as follows. To a suspension of1-methyl-4-(4-nitro-phenyl)-piperazine (2 g, 9.02 mmol) in absoluteethanol (30 mL) was added 10% Pd-C (250 mg). The resultant mixture wasstirred under an atmosphere of H₂ for 5 hours, then filtered through apad of Celite. The filtrate was concentrated under reduced pressure toafford 1.7 g (99% yield) of a brown solid, which was used withoutfurther purification.

[0563]¹H NMR (CDCl₃): δ 6.81 (2H, d, J=8.8 Hz), 6.62 (2H, d, J=8.8 Hz),3.42 (2H, bs), 3.15 (4H, t, J=5.0 Hz), 2.68 (4H, t, J=5.0 Hz), 2.40 (3H,s).

[0564] 1-(4-Isothiocyanato-phenyl)-4-methyl-piperazine, which has thestructural formula

[0565] was prepared in a manner analogous to1-(4-isothio-cyanato-phenyl)-1H-imidazole for Example C(41).4-(4-Methyl-piperazin-1-yl)-aniline provided 1.7 g (83% yield) of acream-colored solid, mp 118-120° C. (lit. 120-122° C., Galstuckova etal., J. Org. Chem. USSR (Engl. Transl), vol. 5 (1969), pp. 1121-1124),which was used without further purification. IR spectrum matched thatreported by Martvon et al., Chem. Zvesti, vol. 27 (1973), pp. 808-810.

[0566]¹H NMR (CDCl₃): δ 7.20 (2H, d, J=9.0 Hz), 6.82 (2H, d, J=9.0 Hz),3.20 (4H, dd, J=5.0, 4.7 Hz), 2.52 (4H, dd, J=5.0, 4.7 Hz), 2.24 (3H,s).

[0567] Anal. Calcd. for C₁₂H₁₅N₃S: C, 61.77; H, 6.48; N, 18.01; S,13.69. Found: C, 61.51; H, 6.56; N, 17.86; S, 13.69.

[0568] The title compound was prepared in a manner like that describedfor Example C(1). 1-(4-Isothiocyanato-phenyl)-4-methyl-piperazine and4-bromoacetyl-3,5-dichloro-pyridine (from Example C(63)) gave a crudesolid, which after recrystallization with EtOH/H₂O, provided a 40 mg(23% yield) of a pale brown solid, mp 150-151° C.

[0569]¹H NMR (DMSO-d₆): δ 10.78 (1H, s), 8.70 (1H, s), 8.00-8.41 (2H,m), 7.24 (2H, bs), 6.88 (2H, d, J=9.0 Hz), 3.08 (4H, dd, J=5.0, 4.7 Hz),2.40 (4H, dd, J=5.0, 4.7 Hz), 2.20 (3H, s).

[0570] IR (KBr): 3395, 2925, 1618, 1546, 1514, 1426, 1240 cm⁻¹.

[0571] HRFABMS: Calcd. for C₂₀H₂₁Cl₂N₆OS (MH⁺): 463.0875. Found:563.0861.

[0572] Anal. Calcd. for C₂₀H₂₀N₆OSCl₂.0.6H₂O.0.1EtOH.0.05CHCl₃: C,50.20; H, 5.06; N, 16.22; S, 6.19, Cl, 14.71. Found: C, 50.34; H, 5.11;N, 16.53; S, 6.43; Cl, 14.74.

Example C(71){4-Amino-2-[4-(4-methyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(2,6-dichloro-phenyl)-methanone

[0573]

[0574] The title compound was prepared in a manner like that describedfor Example C(1). 1-(4-Isothiocyanato-phenyl)-4-methyl-piperazine (fromExample C(70)) and 2-bromo-2′,6′-dichloro-acetophenone (from ExampleC(52)) gave, after recrystallization with H₂O/EtOH/CH₂Cl₂, 2.2 g (64%yield) of a yellow solid, mp 160-162° C.

[0575]¹H NMR (DMSO-d₆): δ 10.60 (1H, s), 8.00 (2H, bs), 7.20-7.41 (4H,m), 6.88 (2H, d, J=9.0 Hz), 3.08 (4H, dd, J 5.0, 4.7 Hz), 2.40 (4H, dd,J=5.0, 4.7 Hz), 2.18 (3H, s).

[0576] IR(KBr): 3394, 3164, 2942, 2810, 1610, 1546, 1427, 1242 cm⁻¹.

[0577] HRFABMS: Calcd. for C₂₁H₂₂Cl₂N₅OS (MH⁺): 462.0922. Found:462.0906.

[0578] Anal. Calcd. for C₂₁H₂₁N₅OSCl₂.0.5H₂O.1EtOH.0.1CH₂Cl₂: C, 52.75;H, 5.40; N, 13.32; S, 6.10, Cl, 14.83. Found: C, 53.06; H, 5.37; N,13.51; S, 6.26; Cl, 14.63.

Example C(72)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(3,5-dibromo-thiophen-2-yl)-methanone

[0579]

[0580] 2-Acetyl-3,5-dibromo-thiophene, which has the structural formula

[0581] was first prepared as follows. To a solution of2,4-dibromothiophene (2.0 g, 8.27 mmol) and acetyl chloride (0.82 mL,11.6 mmol) in ether (3 mL) was added portionwise AlCl₃ (1.5 g, 11.2mmol). After 4 hours, another portion of acetyl chloride and AlCl₃ wereadded, the mixture was refluxed for 1 hour and allowed to cool. Thereaction was carefully quenched with ice and extracted with ether. Theethereal layers were decolorized with activated carbon, dried overMgSO₄, passed through a pad of silica gel, and concentrated to give 1.8g (77% yield) of dark brown oil, which had a ¹H NMR spectrum thatmatched previously described, see del Agua et al, J. Heterocycl. Chem.,vol. 18 (1981), pp. 1345-1347, and was used without furthercharacterization.

[0582] 2-Bromoacetyl-3,5-dibromo-thiophene, which has the structuralformula

[0583] was next prepared in a manner analogous to2-bromo-2′-iodo-acetophenone, see Example C(12).2-Acetyl-3,5-dibromo-thiophene (220 mg, 0.77 mmol) provided 295 mg ofdark brown solid, which was used without further purification.

[0584]¹H NMR (CDCl₃): δ 7.13 (1H, s), 4.54 (2H, s).

[0585] Finally, the title compound was prepared in a manner analogous tothat used in Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boevet al., Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822)and 2-bromoacetyl-3,5-dibromo-thiophene provided a dark brown solid in50% yield, mp 261-264° C.

[0586]¹H NMR (DMSO-d₆): δ 12.50 (1H, bs), 10.94 (1H, s), 8.27 (2H, bs),8.21 (1H, s), 7.87 (1H, bs), 7.57 (1H, d, J=8.7 Hz), 7.36 (1H, s), 7.24(1H, d, J=8.7 Hz).

[0587] HRFABMS (MH⁺): Calcd.: 499.8673. Found: 499.8686.

[0588] Anal. Calcd. for C₁₅H₉N₅OS₂Br₂.0.5H₂O: C, 35.45; H, 1.98; N,13.78; S, 12.62; Br, 31.45. Found: C, 35.37; H, 1.73; N, 13.52; S,12.75; Br, 31.25.

Example C(73)4-[4-Amino-5-(3,5-dibromo-thiophene-2-carbonyl)-thiazol-2-ylamino]-benzenesulfonamide

[0589]

[0590] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-tsothiocyanato-benzenesulfonamide and2-bromoacetyl-3,5-dibromo-thiophene (from Example C(72)) provided ayellow powder in 41% yield, mp 254-255° C.

[0591]¹H NMR (DMSO-d₆): δ 11.24 (1H, s), 8.31 (2H, bs), 7.77 (4H, s),7.40 (1H, s), 7.28 (2H, s).

[0592] FABMS (MH⁺): 536/538/540.

[0593] Anal. Calcd. for C₁₄H₁₀N₄O₃S₃Br₂: C, 31.24; H, 1.87; N, 10.41; S,17.87; Br, 29.69. Found: C, 31.08; H, 1.90; N, 10.16; S, 17.69; Br,29.96.

Example C(74)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(1,5-dimethyl-1H-imidazol-4-yl)-methanone

[0594]

[0595] 1,5-Dimethyl-1H-imidazole-4-carboxylic acid, which has thestructural formula

[0596] was first made as follows. A fresh solution of NaOH (3.86 g, 96.5mmol) in water (20 mL) was added to a solution of ethyl1,5-dimethyl-1H-imidazole-4-carboxylate (5.39 g, 32.0 mmol; Ohno et al,Chem. Pharm. Bull., vol. 42 (1994), pp. 1463-1473) in EtOH (20 mL).After 5 hours, the mixture was cooled to 0° C., and acidified with 38%HCl to pH 3-4. The resultant white solid was filtered off, washed withsmall amount of cold EtOH:H₂O (1:1), and dried under high vacuum to give3.51 g (78%) of white solid, which was used without furtherpurification.

[0597]¹H NMR (D₂O): δ 8.49 (1H, s), 3.73 (3H, s), 2.46 (3H, s).

[0598] Anal. Calcd. for C₆H₈N₂O₂: C, 51.42; H, 5.75; N, 19.99. Found: C,51.52; H, 5.78; N, 19.98.

[0599] 1,5-Dimethyl-1H-imidazole-4-carboxylic acidN-methoxy-N-methyl-amide, which has the structural formula

[0600] was next prepared as follows. To a mixture of1,5-dimethyl-1H-imidazole-4-carboxylic acid (2.01 g, 14.4 mmol) in DMF(20 mL) was addedO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HATU; 6.00 g, 15.8 mmol) and diisopropylethylamine(7.5 mL, 43 mmol). After 5 min., to the resultant clear solution wasadded N,O-dimethylhydroxylamine hydrochloride (1.54 g, 15.79 mmol).After 1 hour, the resultant yellow solution was partitioned betweenCHCl₃ and water. The separated organic layer was washed with water andbrine, dried over K₂CO₃, concentrated, and dried under high vacuum toprovide 1.88 g (72% yield) of light brown solid, which was used withoutfurther purification.

[0601]¹H NMR (CDCl₃): δ 7.36 (1H, s), 3.81 (3H, s), 3.56 (3H, s), 3.47(3H, s), 2.45 (3H, s).

[0602] 1-(1,5-Dimethyl-1H-imidazol-4-yl)-ethanone, which has thestructural formula

[0603] was prepared as follows. To a solution of crude1,5-dimethyl-1H-imidazole-4-carboxylic acid N-methoxy-N-methyl-amide(1.69 g, 9.21 mmol) in THF (55 mL) at −78° C. was added dropwise 1.4 MCH₃MgBr in ether (8.55 mL, 12.0 mmol). The mixture was allowed to warmto ambient temperature over one hour, then quenched with 1N HCl,basified to pH 9 with 1N NaOH, concentrated under reduced pressure toremove the THF, and extracted with EtOAc (200 mL). The organic layer wasseparated, dried over K₂CO₃, and evaporated to furnish 1.2 g (94% yield)of yellow solid, which was used without further purification.

[0604]¹H NMR (CDCl₃): δ 7.35 (1H, s), 3.57 (3H, s), 2.55 (3H, s), 2.53(3H, s).

[0605] 2-Bromo-1-(1,5-dimethyl-1H-imidazol-4-yl)-ethanone, which has thestructural formula

[0606] was next prepared as follows. To1-(1,5-dimethyl-1H-imidazol-4-yl)-ethanone (464 mg, 3.36 mmol) in HOAc(8.5 mL) at 0° C. was added dropwise bromine (173 μl, 3.36 mmol). After36 hours at ambient temperature, crude2-bromo-1-(1,5-dimethyl-1H-imidazol-4-yl)-ethanone hydrobromide salt wasfiltered off as a brown solid, which was successively washed with aminimal amount of water and ether, dissolved in CHCl₃, cooled to 0° C.,treated with NaHCO₃, and concentrated under reduced pressure below 40°C. to obtain 719 mg (99% yield) of yellow oil, which was used withoutfurther purification.

[0607]¹H NMR (DMSO-d₆): δ 8.40 (1H, s), 4.68 (2H, s), 3.66 (3H, s), 2.67(3H, s).

[0608] The title compound was finally prepared in a manner analogous tothat used in Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boevet al., Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822)and 2-bromo-1-(1,5-dimethyl-1H-imidazol-4-yl)-ethanone provided a darkbrown solid in 15% yield, mp _(275.5)-277.0° C.

[0609]¹H NMR (DMSO-d₆): δ 12.42 (11H, s), 10.42 (1H, s), 8.16 (1H, s),7.94 (1H, bs), 7.61-7.30 (2H, m), 7.26 (1H, dd, J=8.4, 1.9 Hz), 3.54(3H, s), 2.51 (3H, s).

[0610] HRFABMS (MH⁺): Calcd.: 354.1137. Found: 354.1132.

[0611] Anal. Calcd. for C₁₆H₁₅N₇OS.0.5H₂O.0.8CH₃OH: C, 52.00; H, 4.99;N, 25.27; S, 8.26. Found: C, 52.27; H, 4.81; N, 25.06; S, 8.12.

Example C(75)[4-Amino-2-(4-morpholin-4-yl-phenylamino)-thiazol-5-yl]-(2,6-dichloro-3-nitro-phenyl)-methanone

[0612]

[0613] 2-Bromo-2′,6′-dichloro-3′-nitro-acetophenone, which has thestructural formula

[0614] was first prepared as follows. To a solution of2′,6′-dichloro-3′-nitro-acetophenone (1.3 g, 5.6 mmol; Breslin, et al.,J. Med. Chem., vol. 38 (1995), pp. 771-793) in glacial acetic acid (5mL) at ambient temperature was added bromine (352 μL, 6.83 mmol). Theresulting mixture was heated to 80° C. for 1 hour, allowed to cool, anddiluted with ether. The organic layer was washed with ice-cold H₂O (25mL), sat aq. NaHCO₃ (3×25 mL), and brine (25 mL), dried over MgSO₄, andconcentrated under reduced pressure to give 1.7 g (97% in crude yield)of a yellow oil, which was used without further purification.

[0615]¹H NMR (CDCl₃): δ 7.98 (1H, d, J=8.7 Hz), 7.38 (1H, d, J=8.7 Hz),4.40 (2H, s).

[0616] The title compound was prepared in a manner like that describedfor Example C(1). 2-Bromo-2′,6′-dichloro-3′-nitro-acetophenone and4-(4-isothiocyanato-phenyl)-morpholine (from Example C(54)) gave a crudesolid, which after purification by flash column chromatography withhexane/EtOAc (70:30) as eluant, provided a dark-brown foam in 52% yield,mp 170-172° C.

[0617]¹H NMR (DMSO-d₆): δ 10.70 (1H, s), 8.30 (1H, s), 8.10 (1H, d,J=9.0 Hz), 7.90 (1H, d, J=8.7 Hz), 7.20-7.30 (2H, m), 6.90 (2H, d, J=9.0Hz), 3.70 (4H, dd, J=5.0, 4.7 Hz), 3.06 (4H, dd, J=5.0, 4.7 Hz).

[0618] IR (KBr): 3289, 2966, 2848, 1634, 1542, 1425, 1343, 1225, 1108cm⁻¹.

[0619] HRFABMS: Calcd. for C₂₀H₁₇Cl₂N₅O₄SNa (M+Na⁺): 516.0276. Found:516.0258.

[0620] Anal. Calcd. for C₂₀H₁₇Cl₂N₅O₄S 0.35CHCl₃: C, 45.59; H, 3.26; N,13.06; S, 5.98, Cl, 20.07. Found: C, 45.33; H, 3.37; N, 12.96; S, 5.93;Cl, 20.27.

Example C(76)4-[4-Amino-5-(1,5-dimethyl-1H-imidazole-4-carbonyl)-thiazol-2-ylamino]-benzenesulfonamide

[0621]

[0622] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Isothiocyanato-benzenesulfonamide and5-bromoacetyl-1,5-dimethyl-1H-imidazole (from Example C(74)) provided ayellow solid in 8% yield, mp 293-294° C.

[0623]¹H NMR (DMSO-d₆): δ 10.80 (1H, s), 7.81 (2H, d, J=9.0 Hz), 7.75(2H, d, J=9.0 Hz), 7.62 (1H, s), 7.24 (2H, s), 3.56 (3H, s), 2.52 (3H,s).

[0624] HRFABMS (M+Na⁺): Calcd.: 415.0623. Found: 415.0609.

[0625] Anal. Calcd. for C₁₅H₁₆N₆O₃S₂.1.0 CH₃OH.1.0CHCl₃: C, 42.53; H,4.45; N, 18.26; S, 13.93. Found: C, 42.57; H, 4.41; N, 18.18; S, 14.07.

Example C(77)[4-Amino-2-(4-morpholin-4-yl-phenylamino)-thiazol-5-yl]-(1,5-dimethyl-1H-imidazol-4-yl)-methanone

[0626]

[0627] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-(4-Isothiocyanato-phenyl)-morpholine (fromExample C(54)) and 5-bromoacetyl-1,5-dimethyl-1H-imidazole (from ExampleC(74)) provided a yellow solid in 12% yield, mp>300° C.

[0628]¹H NMR (DMSO-d₆): δ 10.21 (1H, s), 7.57 (1H, s), 7.42 (2H, d,J=8.8 Hz), 6.94 (2H, d, J=8.8 Hz), 3.72 (4H, t, J=4.7 Hz), 3.54 (3H, s),3.06 (4H, t, J=4.7 Hz), 2.50 (3H, s).

[0629] HRFABMS (M+): Calcd.: 398.1525. Found: 398.1516.

[0630] Anal. Calcd. for C₁₉H₂₂N₆O₂S.0.2 CH₃OH.0.2CHCl₃: C, 54.34; H,5.41; N, 19.60; S, 7.48. Found: C, 54.63; H, 5.27; N, 19.56; S, 7.47.

Example C(78)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(3-methyl-5-nitro-thiophen-2-yl)-methanone

[0631]

[0632] 2-Acetyl-3-methyl-5-nitro-thiophene, which has the structuralformula

[0633] was first prepared as follows. 2-Bromo-3-methyl-5-nitro-thiophene(5.17 g, 23.3 mmol; Spinelli et al, J. Chem. Soc. Perkin Trans. 2,(1975), pp. 620-622), tributyl (1-ethoxyvinyl)tin(IV) (8.65 mL, 25.6mmol), and dichlorobis-(triphenylphosphine) palladium(II) (163 mg, 0.23mmol) in toluene (10.5 mL) was heated under Ar° at 100° C. for 2.5hours. 5% aq HCl (78 mL) was added, and the mixture stirred at 60° C.for 15 min., then partitioned with ether and water. The organic layerwas separated, dried over MgSO₄, and concentrated to a residue that wasdissolved in ether (130 mL). 1,8-Diazabicyclo[5.4.0]undec-7-ene (DBU;2.2 eq) and a 0.1M solution of iodine in ether was added dropwise untilcolor persisted for several seconds. The resultant solution was passedthrough a short column of silica gel and concentrated in vacuo to give3.74 g (87% yield) of yellow solid, which was used without furtherpurification.

[0634]¹H NMR (CDCl₃): δ 7.72 (1H, s), 2.58 (3H, s), 2.57 (3H, s).

[0635] 2-Bromoacetyl-3-methyl-5-nitro-thiophene, which has thestructural formula

[0636] was prepared in a manner analogous to2-bromo-2′-iodo-acetophenone, see Example C(12).2-Acetyl-3-methyl-5-nitro-thiophene (230 mg, 1.24 mmol) provided 330 mgof a cloudy yellow oil, which contained a trace amount of dibrominatedbyproduct by NMR, which was used without further purification.

[0637]¹H NMR (CDCl₃): δ 7.75 (1H, s), 4.28 (2H, s), 2.60 (2H, s).

[0638] The title compound was prepared in a manner analogous to thatused in Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev etal., Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822) and2-bromoacetyl-3-methyl-5-nitro-thiophene provided a yellow solid in 23%yield, mp>300° C.

[0639]¹H NMR (DMSO-d₆): δ 12.50 (1H, d, J=14.3 Hz), 11.01 (1H, bs), 8.40(2H, bs), 8.21 (1H, s), 8.02 (1H, s), 7.63 (1H, bs), 7.52 (1H, bs), 7.36(1H, d, J=11.0 Hz), 2.33 (3H, s).

[0640] HRFABMS (MH⁺): Calcd.: 401.0491. Found: 401.0474.

[0641] Anal. Calcd. for C₁₆H₁₂N₆O₃S₂.0.7H₂O.0.8CH₃OH: C, 46.00; H, 3.81;N, 19.16; S, 14.62. Found: C, 45.92; H, 3.50; N, 19.096; S, 14.59.

Example C(79)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(2,6-difluoro-phenyl)-methanone

[0642]

[0643] 2-Bromo-2′,6′-difluoro-acetophenone, which has the structuralformula

[0644] was first prepared in a manner analogous to2-bromo-2′-iodo-acetophenone, see Example C(12).2′,6′-difluoroacetophenone (703 mg, 4.5 mmol) provided 1.01 g (96%yield) of light yellow oil, which was used without further purification.

[0645]¹H NMR (CDCl₃): δ 7.56-7.42 (1H, m), 7.07-6.98 (2H, m), 4.38 (2H,s).

[0646] The title compound was prepared in a manner analogous to thatused in Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev etal., Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822) and2-bromo-2′,6′-difluoro-acetophenone provided yellow crystals in 78%yield, mp 194-200° C.

[0647]¹H NMR (DMSO-d₆): δ 12.45 (1H, s), 10.86 (1H, s), 8.19 (1H, s),8.16 (2H, bs), 7.80 (1H, bs), 7.59-7.44 (2H, m), 7.22-7.11(3H, m).

[0648] HRFABMS (MH⁺): Calcd.: 372.0731. Found: 372.0725.

[0649] Anal. Calcd. for C₁₇H₁₁N₅OSF₂.0.5H₂O: C, 53.68; H, 3.18; N,18.41; S, 8.43. Found: C, 53.73; H, 3.14; N, 18.32; S, 8.53.

Example C(80){4-Amino-2-[4-(4-methyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(2,6-difluoro-phenyl)-methanone

[0650]

[0651] The title compound was prepared in a manner analogous to thatused in Example C(1). 1-(4-Isothiocyanato-phenyl)-4-methyl-piperazine(from Example C(70)) and 2-bromo-2′,6′-difluoro-acetophenone (fromExample C(79)) provided a yellow solid in 71% yield, mp 168-70° C.

[0652]¹H NMR (DMSO-d₆): δ 10.62 (1H, s), 8.11 (2H, bs), 7.54-7.43 (1H,m), 7.28 (2H, d, J=7.5 Hz), 7.20-7.10 (2H, m), 6.90 (2H, d, J=9.0 Hz),3.08 (4H, t, J=4.8 Hz), 2.41 (4H, t, J=4.8 Hz), 2.19 (3H, s).

[0653] IR (KBr): 2942, 2809, 1620, 1590, 1546, 1516, 1464, 1429, 1238,1002 cm⁻¹.

[0654] HRFABMS (MH⁺): Calcd.: 430.1513. Found: 430.1502.

[0655] Anal. Calcd. for C₂₁H₂₁N₅OSF₂.0.3H₂O: C, 58.00; H, 5.01; N,16.10; S, 7.37. Found: C, 57.98; H, 4.92; N, 16.08; S, 7.42.

Example C(81)({4-Amino-2-[4-(4-methyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(2,6-dichloro-4-trifluoromethyl-phenyl)-methanone

[0656]

[0657] The title compound was prepared in a manner like that describedfor Example C(1). 1-(4-Isothiocyanato-phenyl)-4-methyl-piperazine (fromExample C(70)) and2-bromo-2′,6′-dichloro-4′-trifluoromethyl-acetophenone gave, afterrecrystallization from EtOAc/hexane, yellow needles in 68% yield, mp239-240° C.

[0658]¹H NMR (DMSO-d₆): δ 8.00 (2H, s), 7.28 (2H, bs), 6.92 (2H, d,J=8.7 Hz), 3.10 (4H, dd, J=5.1, 4.7 Hz), 2.42 (4H, dd, J=5.1, 4.8 Hz),2.20 (3H, s).

[0659] IR (KBr): 3377, 3283, 2942, 2813, 1598, 1542, 1513, 1425 cm⁻¹.

[0660] FABMS (M+Na⁺): 552.

[0661] Anal. Calcd. for C₂₂H₂₀Cl₂F₃N₅OS.0.8H₂O.0.7C₆H₁₄: C, 52.00; H,5.23; N, 11.57; S, 5.30, Cl, 11.72. Found: C, 51.94; H, 4.98; N, 11.18;S, 5.20; Cl, 11.48.

Example C(82)N-{3-[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazole-5-carbonyl]-2,4-dichloro-phenyl}-acetamide

[0662]

[0663] 3′-Amino-2′,6′-dichloro-acetophenone, which has the structuralformula

[0664] was first prepared as follows. To a solution of SnCl₂.2H₂O (7.70g, 34.2 mmol) in 6N aq HCl (20 mL) was added2′,6′-dichloro-3′-nitro-acetophenone (4.00 g, 17.1 mmol; Breslin, etal., J. Med. Chem., vol. 38 (1995), pp. 771-793). The resultant mixturewas heated at reflux for 5 hours, allowed to cool, and carefully treatedwith anhydrous Na₂CO₃. The resultant white precipitate was filtered offand washed with CHCl₃. The organic layer was reserved and the aqueouslayer was extracted with CHCl₃ (3×50 mL). The combined organic layerswere dried over MgSO₄, filtered, and concentrated in vacuo to give ablack oil, which was purified via flash column chromatography withEtOAc:hexane (20:80) as eluant. In this manner, 2.6 g (75% yield) of apale brown oil was obtained and used without further purification.

[0665]¹H NMR (CDCl₃): δ 7.08 (1H, d, J=8.7 Hz), 6.70 (1H, d, J=8.7 Hz),4.12 (2H, bs), 2.56 (3H, s).

[0666] N-(3-Acetyl-2,4-dichloro-phenyl)-acetamide, which has thestructural formula,

[0667] was next prepared as follows. To a solution of3′-amino-2′,6′-dichloro-acetophenone (2.40 g, 11.8 mmol) in glacialacetic acid (25 mL) was added acetic anhydride (5.56 mL, 58.8 mmol). Theresultant mixture was heated at reflux for 2 hours, allowed to cool, anddiluted with ether (100 mL). The organic layer was washed with H₂O (2×50mL), dried over MgSO₄, concentrated in vacuo, and azeotroped withn-heptane to give 2.3 g of a pale white solid, which was used withoutfurther purification.

[0668]¹H NMR (CDCl₃): δ 8.38 (1H, d, J=9.1 Hz), 7.62 (1H, bs), 7.34 (1H,d, J=9.0 Hz), 2.60 (3H, s), 2.22 (3H, s).

[0669] N-(3-Bromoacetyl-2,4-dichloro-phenyl)-acetamide, which has thestructural formula,

[0670] was prepared in a manner analogous to2-bromo-2′,6′-dichloro-3′-nitro-acetophenone for Example C(75).N-(3-Acetyl-2,4-dichloro-phenyl)-acetamide gave a pale brown oil in 100%crude yield, which was used without further purification.

[0671]¹H NMR (CDCl₃): δ 8.48 (1H, d, J=8.7 Hz), 7.60 (1H, bs), 7.38 (1H,d, J=9.0 Hz), 4.40 (2H, s), 2.2 (3H, s).

[0672] The title compound was prepared in a manner like that describedfor Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev et al.,Pharm. Chem. J. (Engl. Transl.), vol. 24 (1990), pp. 818-822) andN-(3-bromoacetyl-2,4-dichloro-phenyl)-acetamide gave a product which waspurified via flash column chromatography with a stepwise gradient ofMeOH:CH₂Cl₂ (10:90) to HOAc:MeOH:CH₂Cl₂ (1:10:90) to provide a yellowfoam in 56% yield, that decomposed above 200° C.

[0673]¹H NMR (DMSO-d₆): δ 9.90 (1H, s), 8.20 (1H, s), 7.84-7.96 (1H, m),7.68 (1H, d, J=7.4 Hz), 7.58 (1H, d, J=8.8 Hz), 7.24 (1H, d, J=8.4 Hz),2.20 (3H, s).

[0674] IR(KBr): 3295, 1625, 1525, 1425 cm⁻¹.

[0675] HRFABMS. Calcd. (MH⁺): 461.0354. Found: 461.0344.

[0676] Anal. Calcd. for C₁₉H₁₅Cl₂N₆O₂S.H₂O.3HOAc: C, 45.53; H, 4.28; N,12.74; S, 4.86, Cl, 10.75. Found: C, 45.93H, 4.08; N, 12.49; S, 4.83;Cl, 10.45.

Example C(83)[4-Amino-2-(4-morpholin-4-yl-phenylamino)-thiazol-5-yl]-(3-methyl-biphenyl-2-yl)-methanone

[0677]

[0678] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-(4-Isothiocyanato-phenyl)-morpholine (fromExample C(54)) and 2-bromoacetyl-3-methyl-biphenyl (from Example C(67))provided a yellow solid in 29% yield, mp 125-35° C.

[0679]¹H NMR (DMSO-d₆): δ 10.40 (1H, s), 7.86 (2H, s), 7.42-7.24 (9H,m), 7.19 (1H, d, J=7.5 Hz), 6.93 (2H, d, J=8.7 Hz), 3.73 (4H, t, J=4.4Hz), 3.07 (4H, t, J=4.4 Hz), 2.26 (3H, s).

[0680] HRFABMS (M⁺): Calcd.: 471.1855. Found: 471.1839.

[0681] Anal. Calcd. for C₂₇H₂₆N₄O₂S.1.0CF₃CO₂H: C, 59.58; H, 4.66; N,9.58; S, 5.48. Found: C, 59.41; H, 5.01; N, 9.26; S, 5.18.

Example C(84)[4-Amino-2-(4-morpholin-4-yl-phenylamino)-thiazol-5-yl]-(2-bromo-6-methyl-phenyl)-methanone

[0682]

[0683] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-(4-Isothiocyanato-phenyl)-morpholine (fromExample C(54)) and 2,2′-dibromo-6′-methyl-acetophenone (from ExampleC(66)) provided a crude solid, which was triturated with MeOH/CHCl₃ tofurnish a yellow solid in 22% yield, mp 105-125° C.

[0684]¹H NMR (DMSO-d₆): δ 10.57 (1H, s), 8.01 (2H, bs), 7.46 (1H, d,J=7.5 Hz), 7.39-7.18 (4H, m), 6.96 (2H, d, J=8.7 Hz), 3.74 (4H, t, J=4.7Hz), 3.09 (4H, t, J=4.7 Hz), 2.20 (3H, s).

[0685] HRFABMS (MH⁺): Calcd.: 73.0647/475. Found: 473.0657/475.

[0686] Anal. Calcd. for C₂₁H₂₁N₄O₂SBr.0.7 MeOH.0.6CHCl₃: C, 47.60; H,4.37; N, 9.98; S, 5.71. Found: C, 47.95; H, 4.05; N, 9.77; S, 5.51.

Example C(85)4-[4-Amino-5-(2,6-difluoro-benzoyl)-thiazol-2-ylamino]-benzenesulfonamide

[0687]

[0688] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Isothiocyanato-benzenesulfonamide and2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided lightyellow crystals in 69% yield, mp 258-260° C.

[0689]¹H NMR (DMSO-d₆): δ 11.20 (1H, s), 8.20 (2H, bs), 7.79 (2H, d,J=9.0 Hz), 7.74 (2H; d, J=9.0 Hz), 7.61-7.49 (1H, m), 7.26 (2H, s), 7.22(1H, d, J=7.9 Hz), 7.19 (1H, d, J=8.0 Hz).

[0690] IR(KBr): 3310, 1622, 1599, 1547, 1525, 1467, 1425, 1410, 1318,1156 cm¹.

[0691] HRFABMS (MH⁺): Calcd.: 411.0397. Found: 411.0410.

[0692] Anal. Calcd. for C₁₆H₁₂N₄O₃S₂F₂.0.7CH₃OH: C, 46.34; H, 3.45; N,12.94; S, 14.82. Found: C, 46.19; H, 3.12; N, 12.83; S, 14.94.

Example C(86)N-(3-{4-Amino-2-[4-(4-methyl-piperazin-1-yl)-phenylamino]-thiazole-5-carbonyl}-2,4-dichloro-phenyl)-acetamide

[0693]

[0694] The title compound was prepared in a manner like that describedfor Example C(1). 1-(4-Isothiocyanato-phenyl)-4-methyl-piperazine (fromExample C(70)) and N-(2-bromoacetyl-3-chloro-phenyl)-acetamide (fromExample C(82)) gave, after recrystallization with EtOH/CHCl₃, 60 mg (13%yield) of a yellow solid, mp 195-197° C.

[0695]¹H NMR (DMSO-d₆): δ 10.62 (1H, s), 9.62 (1H, s), 7.90 (1H, bs),7.78 (1H, dd, J=8.9, 4.4 Hz), 7.47 (1H, d, J=8.8 Hz), 7.30 (2H, bs),6.92 (2H, d, J=9.1 Hz), 3.08 (4H, dd, J=5.1, 4.6 Hz), 2.42 (4H, dd,J=5.1, 4.6 Hz), 2.18 (3H, s), 2.08 (3H, s).

[0696] IR (KBr): 3260, 3025, 2801, 1666, 1613, 1525, 1437, 1382, 1299cm⁻¹.

[0697] HRFABMS. Calcd. (M+Na⁺): 541.0956. Found: 541.0970.

[0698] Anal. Calcd. for C₂₃H₂₄Cl₂N₆O₂S.0.5H₂O.0.4EtOH: C, 52.27; H,5.05; N, 15.37; S, 4.86, Cl, 12.97. Found: C, 52.13; H, 5.09; N, 15.13;S, 5.78; Cl, 12.96.

Example C(87){4-Amino-2-[4-(4-methyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-3-methyl-thiophen-2-yl-methanone

[0699]

[0700] The title compound was prepared in a manner like that describedfor Example C(1). 1-(4-Isothiocyanato-phenyl)-4-methyl-piperazine (fromExample C(70)) and 2-bromoacetyl-3-methyl-thiophene (from Example C(19))gave, after recrystallization with EtOH/CHCl₃, a dark yellow solid in75% yield, mp 237.0-237.5° C.

[0701]¹H NMR (DMSO-d₆): δ 10.50 (1H, s), 8.10 (1H, bs), 7.56 (1H, d,J=5.0 Hz), 7.38 (2H, d, J=8.8 Hz), 6.96 (3H, m), 3.10 (4H, dd, J=5.1,4.7 Hz), 2.45 (4H, dd, J=4.9, 4.7 Hz), 2.38 (3H, s), 2.24 (3H, s).

[0702] IR (KBr): 3484, 3319, 2943, 2809, 1593, 1546, 1414 cm⁻¹.

[0703] HRFABMS. Calcd. (MH⁺): 414.1422. Found: 414.1408.

[0704] Anal. Calcd: C₂₀H₂₃N₅OS₂.3H₂O: C, 57.34; H, 5.68; N, 16.72; S,15.31. Found: C, 57.01; H, 5.72; N, 16.41; S, 15.34.

Example C(88)trans-3RS-Amino-4RS-{4-[4-amino-5-(3,5-dichloropyridine-4-carbonyl)-thiazol-2-ylamino]-benzoyl}-dihydro-furan-2-one

[0705]

[0706] The title compound was prepared essentially as described forExample C(1). 4-Isothiocyanato-benzoyl-DL-homoserine lactone and4-bromoacetyl-3,5-dichloropyridine (from Example C(63)) gave a productwhich was purified via column chromatography with 10% MeOH/CHCl₃ aseluant to provide an amorphous yellow solid, 203 mg (79%), thatdecomposed above 150° C.

[0707]¹H NMR (DMSO-d₆): δ 11.17 (1H, s), 8.89 (1H, d, J=8.0 Hz), 8.76(2H, s), 7.86 (2H, d, J=8.7 Hz), 7.69 (2H, d, J=8.7 Hz), 4.73 (1H, q,J=9.3 Hz), 4.42 (1H, ddd, J=8.9, 8.7, 1.8 Hz), 4.27 (1H, ddd, J=10.0,8.7, 6.7 Hz).

[0708] HRFABMS. Calcd. for C₂₀H₁₅Cl₂N₅O₄SNa (M+Na⁺): 514.0120. Found:514.0133.

[0709] IR(KBr): 3284, 1774, 1610, 1524, 1459, 1423, 1348. 1306, 1180cm^(−1.)

[0710] Anal. Calcd for C₂₀H₁₅Cl₂N₅O₄S.0.25H₂O.0.6CHCl₃: C, 43.52; H,2.85; N, 12.32; Cl, 23.70; S, 5.64. Found: C, 43.31; H, 2.78; N, 12.46;Cl, 24.07; S, 5.63.

Example C(89)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(2,6-dichloro-3-nitro-phenyl)-methanone

[0711]

[0712] The title compound was prepared in a manner like that describedfor Example C(1). 2-Bromo-2′,6′-dichloro-3′-nitro-acetophenone (fromExample C(75)) and 6-isothiocyanato-1H-benzoimidazole (see Boev et al.,Pharm. Chem. J. (Engl. Transl.), vol. 24 (1990), pp. 818-822) provided,after column chromatography with 1% HOAc/10% MeOH/CH₂Cl₂ as eluant, 26%yield of yellow powder, mp 250-252° C.

[0713]¹H NMR (DMSO-d₆): δ 8.18 (1H, s), 8.00 (2H, d, J=8.7 Hz), 7.80(1H, d, J=8.7 Hz), 7.52 (1H, bd, J=8.1 Hz), 7.24-7.10 (2H, m).

[0714] IR(KBr): 3385, 1607, 1500 cm⁻¹.

[0715] HRFABMS: Calcd. for C₁₇H₉Cl₂N₆O₃S(M−H⁺): 447.9930. Found:447.9930.

[0716] Anal. Calcd. for C₁₇H₁₀Cl₂N₆O₃S.0.1H₂O.1 MeOH.0.7 HOAc.0.1CH₂Cl₂:C, 43.30; H, 3.35; N, 15.54; S, 5.93, Cl, 14.42. Found: C, 43.26; H,3.01; N, 14.74; S, 7.14; Cl, 14.74.

Example C(90){4-Amino-2-[4-(4-methyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(2,5-dimethyl-thiophen-3-yl)-methanone

[0717]

[0718] The title compound was prepared in a manner like that describedfor Example C(1). 3-Bromoacetyl-2,5-dimethyl-thiophene (from ExampleC(50)) and 1-(4-isothiocyanato-phenyl)-4-methyl-piperazine (from ExampleC(70)) gave, after purification via flash column chromatography with5-10% MeOH:CH₂Cl₂ as stepwise gradient eluant, a yellow solid in 70%yield, mp 205-206° C.

[0719]¹H NMR (DMSO-d₆): δ 10.50 (1H, s), 8.00 (2H, bs), 7.48 (2H, d,J=8.7 Hz), 6.95 (2H, d, J=8.7 Hz), 6.80 (1H, s), 3.10 (4H, dd, J=5.0,4.4 Hz), 2.46 (4H, t, J=4.7 Hz), 2.42 (3H, s), 3.38 (3H, s), 2.24 (3H,s).

[0720] IR(KBr): 3154, 2944, 2804, 1609, 1543, 1516, 1420, 1296 cm⁻¹.

[0721] HRFABMS: Calcd. for C₂₁H₂₆N₅OS₂ (MH⁺): 428.1579. Found: 428.1566.

[0722] Anal. Calcd. for C₂₁H₂₅N₅OS₂.0.7H₂O: C, 57.30; H, 6.04; N, 15.91;S, 14.57. Found: C, 57.19; H, 6.06; N, 15.77; S, 14.55.

Example C(91)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(3-amino-4-bromo-2,6-dichloro-phenyl)-methanone

[0723]

[0724] 3′-Amino-4′-bromo-2′,6′-dichloro-acetophenone, which has thestructural formula

[0725] was first prepared as follows.3′-Amino-2′,6′-dichloro-acetophenone (from Example C(82); 2.15 g, 11.3mmol) in HOAc (8.7 mL) was carefully degassed with argon and cooled to0° C., bromine was added, and the reaction mixture was then allowed towarm to ambient temperature. After 0.5 hour, the mixture was dilutedwith ice/water and extracted with ether. The combined ethereal layerswere washed with sat aq. NaHCO₃ and brine, dried over K₂CO₃, andevaporated to afford 2.87 g (90%) of brown solid, which was used withoutfurther purification.

[0726] 3′-Amino-2,4′-dibromo-2′,6′-dichloro-acetophenone, which has thestructural formula

[0727] was prepared in a manner analogous to5-bromoacetyl-4-methyl-1H-imidazole for Example C(40).3′-Amino-4′-bromo-2′,6′-dichloro-acetophenone provided, after columnchromatography with a stepwise gradient of 2.5-5% CH₂Cl₂/hex, 725 mg(22% yield) of white solid, which was used without further purification.Later fractions yielded 33% recovery of starting material.

[0728] The title compound was prepared in a manner analogous to thatused in Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev etal., Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822) and3′-amino-2,4′-dibromo-2′,6′-dichloro-acetophenone provided light yellowcrystals in 34% yield, mp 227-230° C.

[0729]¹H NMR (DMSO-d₆): δ 12.48 (1H, bs), 10.85 (1H, s), 8.22 (1H, s),8.06 (2H, bs), 7.80 (1H, bs), 7.34 (1H, s), 7.58 (1H, d, J=8.5 Hz), 7.22(1H, d, J=8.5 Hz), 5.75 (2H, s).

[0730] FABMS (MH⁺): Calcd.: 498.9333. Found: 498.9312.

[0731] Anal. Calcd. for C₁₇H₁₁N₆OSCl₂Br.0.8H₂O: C, 39.83; H, 2.48; N,16.39; S, 6.26. Found: C, 39.92; H, 2.43; N, 16.26; S, 6.14.

Example C(92)4-[4-Amino-5-(2,5-dichloro-thiophene-3-carbonyl)-thiazol-2-ylamino]-benzenesulfonamide

[0732]

[0733] 3-Bromoacetyl-2,5-dichloro-thiophene, which has the structuralformula

[0734] was prepared in a manner analogous to2-bromo-2′-iodo-acetophenone, see Example C(12):3-Acetyl-2,5-dichlorothiophene (2.0 g, 10.2 mmol) provided 2.9 g (100%yield) of yellow oil, which was used without further purification.

[0735]¹H NMR (CDCl₃): δ 7.25 (1H, s), 4.40 (2H, s).

[0736] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Isothiocyanato-benzenesulfonamide and3-bromoacetyl-2,5-dichloro-thiophene provided a yellow solid in 65%yield, mp 274-276° C.

[0737]¹H NMR (DMSO-d₆): δ 11.20 (1H, s), 8.24 (2H, bs), 7.80 (4H, s),7.33 (1H, s), 7.31 (2H, s).

[0738] FABMS (MH⁺): 449/451.

[0739] Anal. Calcd. for C₁₄H₁₀N₄O₃S₃Cl₂: C, 37.42; H, 2.24; N, 12.47; S,21.41; Cl, 15.78. Found: C, 37.56; H, 2.19; N, 12.39; S, 21.29; Cl,15.71.

Example C(93)[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(2,5-dichloro-thiophen-3-yl)-methanone

[0740]

[0741] The title compound was prepared in a manner analogous to thatused in Example C(1). 6-Isothiocyanato-1H-benzoimidazole (see Boev etal., Pharm. Chem. J. (Engl. Transl)., vol. 24 (1990), pp. 818-822) and3-bromoacetyl-2,5-dichloro-thiophene (from Example C(92)) provided,after precipitation with THF, an amorphous yellow solid in 52% yield,mp>300° C.

[0742]¹H NMR (DMSO-d₆): δ 12.52 (1H, bs), 10.89 (1H, s), 8.26 (1H, s),8.21 (2H, bs), 7.90 (1H, bs), 7.60 (1H, d, J=8.4 Hz), 7.28 (1H, s), 7.27(1H, d, J=8.4 Hz).

[0743] ESIMS(MH⁺): 410/412.

[0744] Anal. Calcd. for C₁₅H₉N₅OS₂Cl₂.0.1HCl.0.6THF: C, 45.71; H, 3.06;N, 15.32; S, 14.03; Cl, 16.28. Found: C, 45.84; H, 2.83; N, 15.01; S,14.27; Cl, 16.00.

Example C(94){4-Amino-2-[4-(4-methyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(3-methyl-5-nitro-thiophen-2-yl)-methanone

[0745]

[0746] The title compound was prepared in a manner analogous to thatused in Example C(1). 1-(4-Isothiocyanato-phenyl)-4-methyl-piperazine(from Example C(70)) and 2-bromoacetyl-3-methyl-5-nitro-thiophene (fromExample C(78)) afforded, after precipitation with aqueous EtOH, anamorphous dark brown solid in 64% yield.

[0747]¹H NMR (DMSO-d₆): δ 10.88 (1H, s), 8.38 (2H, bs), 8.04 (1H, s),7.38 (2H, d, J=9.0 Hz), 6.98 (2H, d, J=9.0 Hz), 3.35 (4H, bs), 3.15 (4H,bs), 2.34 (3H, s), 2.28 (3H, s).

[0748] HRFABMS (MH⁺): Calcd.: 459.1273. Found: 459.1259.

[0749] Anal. Calcd. for C₂₀H₂₂N₆O₃S₂.0.8H₂O.0.2EtOH: C, 50.81; H, 5.18;N, 17.43; S, 13.30. Found: C, 50.94; H, 4.98; N, 17.13; S, 13.55.

Example C(95)4-[4-Amino-5-(3-methyl-5-nitro-thiophene-2-carbonyl)-thiazol-2-ylamino]-benzenesulfonamide

[0750]

[0751] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Isothiocyanato-benzenesulfonamide and2-bromoacetyl-3-methyl-5-nitro-thiophene (from Example C(78)) provided adark brown in 38% yield, mp 268-269° C.

[0752]¹H NMR (DMSO-d₆): δ 11.31 (1H, s), 8.46 (2H, bs), 8.08 (1H, s),7.81 (4H, s), 7.32 (2H, s), 2.38 (3H, s).

[0753] Anal. Calcd. for C₁₅H₁₃N₅O₅S₃: C, 40.99; H, 2.98; N, 15.94; S,21.89. Found: C, 41.11; H, 2.95; N, 15.66; S, 21.70.

Example C(96)(3-Amino-4-bromo-2,6-dichloro-phenyl)-{4-amino-2-[4-(4-methylpiperazin-1-yl)-phenylamino]-thiazol-5-yl}-methanone

[0754]

[0755] The title compound was prepared in a manner analogous to thatused in Example C(1). 1-(4-Isothiocyanato-phenyl)-4-methyl-piperazine(from Example C(70)) and3′-amino-2,4′-dibromo-2′,6′-dichloro-acetophenone (from Example C(91))provided, after recrystallization from EtOH, a yellow powder in 43%yield, mp 180-182° C.

[0756]¹H NMR (DMSO-d₆): δ 10.61 (1H, s), 8.01 (2H, bs), 7.59 (1H, s),7.28 (2H, d, J=8.7 Hz), 6.94 (2H, d, J=8.7 Hz), 5.74 (2H, s), 3.11 (4H,bs), 2.45 (4H, bs), 2.23 (3H, s).

[0757] HRFABMS (MH⁺): Calcd.: 555.0136/557/559. Found: 555.0122/557/559.

[0758] Anal. Calcd. for C₂₁H₂₁N₆OSCl₂Br.0.7H₂O.0.6EtOH: C, 44.70; H,4.39; N, 14.09; S, 5.38. Found: C, 44.84; H. 4.18; N, 13.95; S, 5.27.

Example C(97)2-[4-(1-Acetyl-piperazin-4-yl)-phenylamino]-4-amino-thiazol-5-yl-(2,6-dichlorophenyl)-methanone

[0759]

[0760] 1-Acetyl-4-(4-nitro-phenyl)-piperazine, which has the structuralformula

[0761] was first prepared in a manner analogous toN-(3-acetyl-2,4-dichloro-phenyl)-acetamide for Example C(82).1-(4-Nitro-phenyl)-piperazine gave a yellow solid in 83% yield, whichmatched previously reported material by ¹H NMR (Katz et al., J. Amer.Chem. Soc., vol. 111 (1989), pp. 7554-7557).

[0762] 1-Acetyl-4-(4-amino-phenyl)-piperazine, which has the structuralformula

[0763] was prepared in a manner analogous to4-(4-methyl-piperazin-1-yl)-aniline for Example C(82).1-Acetyl-4-(4-nitro-phenyl)-piperazine gave a pale white powder in 100%crude yield, which was used without any further purification.

[0764]¹H NMR (CDCl₃): δ 6.85 (2H, d, J=8.7 Hz), 6.98 (2H, d, J=8.7 Hz),3.78 (2H, dd, J=5.3, 5.0 Hz), 3.62 (2H, t, J=5.3, 5.0 Hz), 3.62 (2H, dd,J=5.3, 5.0 Hz), 2.98-3.10 (4H, m), 2.18 (3H, s).

[0765] 1-Acetyl-4-(4-isothiocyanato-phenyl)-piperazine, which has thestructural formula

[0766] was prepared in a manner analogous to1-(4-isothiocyanato-phenyl)-1H-imidazole for Example C(41).1-Acetyl-4-(4-amino-phenyl)-piperazine provided a cream-colored powderin 88% yield.

[0767]¹H NMR (CDCl₃): δ 7.18 (2H, d, J=9.0 Hz), 6.82 (2H, d, J=9.0 Hz),3.78 (2H, dd, J=5.1, 5.3 Hz), 3.64 (2H, dd, J=4.9, 5.3 Hz), 3.16-3.27(4H, m), 2.10 (3H, s).

[0768] The title compound was prepared in a manner like that describedfor Example C(1). 2-Bromo-2′,6′-dichloro-acetophenone (from ExampleC(52)) and 1-acetyl-4-(4-isothiocyanato-phenyl)-piperazine gave a crudeproduct that precipitated from hexanes to provide a cream solid in 37%yield, mp 265-267° C.

[0769]¹H NMR (DMSO-d₆): δ 10.60 (1H, bs), 8.02 (2H, bs), 7.50 (2H, d,J=1.9 Hz), 7.42 (1H, m), 7.38 (2H, bs), 6.98 (2H, d, J=9.0 Hz), 3.60(4H, s), 3.20-3.10 (4H, m), 2.00 (3H, s).

[0770] IR(KBr): 3377, 3166, 1601, 1542, 11425 cm⁻¹.

[0771] HRFABMS: Calcd. for C₂₂H₂₂Cl₂N₅O₂S (MH⁺): 490.0871. Found:490.0858.

[0772] Anal. Calcd. for C₂₂H₂₁Cl₂N₅O₂S.0.16H₂O.0.1C₆H₁₄: C, 54.08; H,4.56; Cl, 14.13; N, 13.95; S, 6.39. Found: C, 53.88; H, 4.32; Cl, 14.46;N, 14.28; S, 6.54.

Example C(98)2-[4-(1-Acetyl-piperazine-4-yl)-phenylamino]-4-amino-thiazol-5-yl-(3-methyl-thiophen-2-yl)-methanone

[0773]

[0774] The title compound was prepared in a manner like that describedfor Example C(1). 1-Acetyl-4-(4-isothiocyanato-phenyl)-piperazine (fromExample C(97)) and 2-bromoacetyl-3-methyl-thiophene (from Example C(l9)) provided a yellow solid in 37% yield, mp 290-292° C.

[0775]¹H NMR (DMSO-d₆): δ 10.60 (1H, bs), 8.10 (2H, bs), 7.48 (1H, d,J=5.0 Hz), 7.40 (2H, d, J=8.7 Hz), 6.96-7.04 (2H, m), 3.60 (4H, s), 3.18(2H, bs), 3.12 (2H, bs), 2.40 (3H, s), 2.02 (3H, s).

[0776] IR (KBr): 3377, 3166, 1633, 1601, 1542, 1425, 1225 cm⁻¹.

[0777] Anal. Calcd. for C₂₁H₂₃N₅O₂S.1H₂O: C, 56.89; H, 5.27; N, 15.80;S, 14.46. Found: C, 56.98; H, 5.27; N, 15.72; S, 14.35.

Example C(99)4-[4-Amino-5-(2-fluoro-6-trifluoromethyl-benzoyl)-thiazol-2-ylamino]-benzenesulfonamide

[0778]

[0779] 2-Bromo-2′-fluoro-6′-trifluoromethyl-acetophenone, which has thestructural formula

[0780] was prepared in a manner analogous to2-bromo-2′-iodo-acetophenone, see Example C(12).2′-Fluoro-6′-(trifluoromethyl)-acetophenone (745 mg, 3.61 mmol) provided1.05 g of yellow oil, which was used without further purification.

[0781]¹H NMR (CDCl₃): δ 7.69-7.52 (2H, m), 7.44-7.35 (1H, m), 4.42 (3H,s).

[0782] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Isothiocyanato-benzenesulfonamide and crude2-bromo-2′-fluoro-6′-trifluoromethyl-acetophenone provided a lightyellow solid in 21% yield, mp 290-292° C.

[0783]¹H NMR (DMSO-d₆): δ 11.15 (1H, s), 8.20 (2H, bs), 7.83-7.68 (7H,m), 7.31 (2H, s).

[0784] Anal. Calcd. for C₁₇H₁₂N₄O₃S₂F₄: C, 44.35; H, 2.63; N, 12.17; S,13.93. Found: C, 44.42; H, 2.64; N, 12.13; S, 13.94.

Example C(100){4-Amino-2-[4-(4-methyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(2-fluoro-6-trifluoromethyl-phenyl)-methanone

[0785]

[0786] The title compound was prepared in a manner analogous to thatused in Example C(1). 1-(4-Isothiocyanato-phenyl)-4-methyl-piperazine(from Example C(70)) and2-bromo-2′-fluoro-6′-trifluoromethyl-acetophenone (from Example C(99))produced a crude product that recrystallized from EtOH to provide ayellow powder in 74% yield, mp 155-158° C.

[0787]¹H NMR (DMSO-d₆): δ 10.62 (1H, s), 8.06 (2H, bs), 7.72-7.62 (3H,m), 7.10 (2H, d, J=8.7 Hz), 6.93 (2H, d, J=8.7 Hz), 3.11 (4H, bs), 2.45(4H, bs), 2.22 (3H, s).

[0788] HRFABMS (MH⁺): Calcd.: 480.1481. Found: 480.1468.

[0789] Anal. Calcd. for C₂₂H₂₁N₅OSF₄.1.0EtOH: C, 54.84; H, 5.18; N,13.33; S, 6.10. Found: C, 55.11; H, 5.11; N, 13.31; S, 6.00.

Example C(101)4-Amino-2-[4-(1-tert-butoxycarbonyl-piperazine-4-yl)-phenylamino]-thiazol-5-yl-(2,6-difluorophenyl)-methanone

[0790]

[0791] 1-tert-Butoxycarbonyl-4-(4-nitro-phenyl)-piperazine, which hasthe structural formula

[0792] was first prepared as follows. To a suspension of1-(4-nitro-phenyl)-piperazine (2.00 g, 9.65 mmol) in dioxane (30 mL) wasadded diisopropylethylamine (1.48 mL, 10.6 mmol) and di-t-butyldicarbonate (2.10 g, 9.65 mmol). After 12 hours, the mixture was pouredinto H₂O (100 mL) and extracted with EtOAc (2×50 mL). The combinedorganic layers were dried over MgSO₄, filtered, and concentrated invacuo to give a yellow solid, which recrystallized from EtOAc/hexane toafford 2.2 g of yellow needles. This material was used without furtherpurification.

[0793]¹H NMR (CDCl₃): δ 8.20 (2H, d, J=9.3 Hz), 6.82 (2H, d, J=9.3 Hz),3.58-3.64 (4H, m), 3.28-3.44 (4H, m), 1.54 (9H, s).

[0794] 1-(4-Amino-phenyl)-4-tert-butoxycarbonyl-piperazine, which hasthe structural formula

[0795] was prepared in a manner analogous to4-(4-methylpiperazin-1-yl)-aniline for Example C(70).1-tert-Butoxycarbonyl-4-(4-nitro-phenyl)-piperazine furnished a palebrown gel in 100% crude yield, which was used without furtherpurification.

[0796]¹H NMR (CDCl₃): δ 6.84 (2H, d, J=8.7 Hz), 6.67 (2H, d, J=8.8 Hz),3.58 (4H, dd, J=5.1, 5.0 Hz), 2.97 (4H, dd, J=5.2, 4.8 Hz), 1.52 (9H,s).

[0797] 1-tert-Butoxycarbonyl-4-(4-isothiocyanato-phenyl)-piperazine,which has the structural formula

[0798] was prepared in a manner analogous to1-(4-isothiocyanato-phenyl)-1H-imidazole for Example C(41).1-(4-Amino-phenyl)-4-tert-butoxycarbonyl-piperazine affordedcream-colored needles in 87% yield, which were used without furtherpurification.

[0799]¹H NMR (CDCl₃): δ 7.18 (2H, d, J=9.0 Hz), 6.82 (2H, d, J=9.0 Hz),3.64 (4H, t, J=5.3 Hz), 3.24 (4H, t, J=5.3 Hz), 1.54 (9H, s).

[0800] The title compound was prepared in a manner analogous to thatused in Example C(1).1-tert-Butoxycarbonyl-4-(4-isothiocyanato-phenyl)-piperazine and2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) gave a crudeproduct which recrystallized from EtOH to furnish a yellow solid in 67%yield, mp 140-143° C.

[0801]¹H NMR (DMSO-d₆): δ 10.67 (1H, s), 8.13 (2H, bs), 7.59-7.45 (1H,m), 7.35 (2H, d, J=9.0 Hz), 7.23-7.13 (2H, m), 6.96 (2H, d, J=9.0 Hz),3.46 (4H, bs), 3.07 (4H, bs), 1.43 (9H, s).

[0802] HRFABMS (MH⁺): Calcd.: 516.1881. Found: 516.1900.

[0803] Anal. Calcd. for C₂₅H₂₇N₅O₃SF₂.0.8H₂O.0.8EtOH: C, 56.56; H, 5.57;N, 12.99; S, 5.95. Found: C, 56.34; H, 5.54; N, 12.89; S, 5.83.

Example C(102)4-[4-Amino-5-(2,6-difluoro-benzoyl)-thiazol-2-ylamino]-benzamide

[0804]

[0805] 4-Isothiocyanato-benzamide, which has the structural formula

[0806] was first prepared according to a method from McKee et al., J.Am. Chem. Soc., vol. 48 (1946), pp. 2506-2507. To a solution of4-aminobenzamide (5.00 g, 36.7 mmol) in water (60 mL) and 38% aq HCl (15mL) was added thiophosgene (3.08 mL, 40.4 mmol). After approximately 30min, the resultant white precipitate was filtered off, washed withwater, and dried under high vacuum to obtain 4.66 g (78% yield) of whitepowder, which was used without further purification.

[0807]¹H NMR (DMSO-d₆): δ 8.08 (1H, bs), 7.94 (2H, d, J=8.7 Hz), 7.53(2H, d, J=8.7 Hz), 7.51 (1H, bs).

[0808] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Isothiocyanato-benzamide and2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided ayellow solid in 26% yield, mp 297-298° C.

[0809]¹H NMR (DMSO-d₆): δ 11.07 (1H, s), 8.22 (2H, bs), 7.91 (1H, s),7.88 (2H, d, J=8.7 Hz),), 7.66 (2H, d, J=8.7 Hz), 7.62-7.50 (1H, m),7.31 (1H, s), 7.27-7.18 (2H, m).

[0810] Anal. Calcd. for C₁₇H₁₂N₄O₂SF₂: C, 54.54; H, 3.23; N, 14.97; S,8.57. Found: C, 54.27; H, 3.27; N, 14.68; S, 8.35.

Example C(103) tert-Butyl({4-[4-Amino-5-(2,6-difluoro-benzoyl)-thiazol-2-ylamino]-phenyl}-methyl-amino)-acetate

[0811]

[0812] tert-Butyl [methyl-(4-nitro-phenyl)-amino]-acetate, which has thestructural formula

[0813] was first prepared as follows. To a solution of sarcosine t-butylester hydrochloride (2.0 g, 11 mmol) in DMSO (6 mL) was added4-fluoro-nitrobenzene (1.6 g, 11 mmol) and triethylamine (3.4 mL, 24mmol). The resultant mixture was heated at 100° C. for 12 hours. Theresultant yellow suspension was allowed to cool, diluted with H₂O (100mL), and extracted with ether (2×100 mL). The combined organic layerswere dried over MgSO₄, filtered, and concentrated in vacuo to giveyellow needles, which recrystallized from ether/hexane to give 2.0 g ofyellow needles, which were used without further purification.

[0814]¹H NMR (CDCl₃): δ 8.18 (2H, d, J=9.3 Hz), 6.62 (2H, d, J=9.7 Hz),4.08 (2H, s), 3.20 (3H, s), 1.42 (9H, s).

[0815] tert-Butyl [(4-amino-phenyl)-methyl-amino]-acetate, which has thestructural formula

[0816] was prepared in a manner analogous to4-(4-methyl-piperazin-1-yl)-aniline for Example C(70). tert-Butyl[methyl-(4-nitro-phenyl)-amino]-acetate provided a red oil in 95% crudeyield, which was used without further purification.

[0817]¹H NMR (CDCl₃): δ 6.60-6.80 (4H, m), 4.08 (2H, s), 3.20 (2H, bs),3.80 (2H, s), 2.82 (3H, s), 1.42 (9H, s).

[0818] tert-Butyl [(4-isothiocyanato-phenyl)-methyl-amino]-acetate,which has the structural formula

[0819] was prepared in a manner analogous to1-(4-isothiocyanato-phenyl)-1H-imidazole for Example C(41). tert-Butyl[(4-amino-phenyl)-methyl-amino]-acetate furnished a pale brown solid in98% yield, which was used without further purification.

[0820]¹H (CDCl₃): δ 7.10 (2H, d, J=9.1 Hz), 6.52 (2H, d, J=9.1 Hz), 3.90(2H, s), 2.92 (3H, s), 1.30 (9H, s).

[0821] The title compound was prepared in a manner like that describedfor Example C(1). tert-Butyl[(4-isothiocyanato-phenyl)-methyl-amino]-acetate and2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided acream powder in 34% yield, mp 200.0-200.5° C.

[0822]¹H NMR (DMSO-d₆): δ 7.44-7.56 (1H, m), 7.10-7.30 (4H, m), 6.62(2H, d, J=9.0 Hz), 4.08 (2H, s), 2.95 (3H, s), 1.32 (9H, s).

[0823] IR (KBr): 3248, 3142, 2978, 1725, 1619, 1537, 1466, 1231 cm⁻¹.

[0824] Anal. Calcd. for C₂₃H₂₄F₂N₄O₃S: C, 58.22; H, 5.10; N, 11.81; S,6.76. Found: C, 58.27; H, 5.11; Cl, N, 11.53; S, 6.63.

Example C(104)4-Amino-2-[4-(1-tert-butoxycarbonyl-piperazine-4-yl)-phenylamino]-thiazol-5-yl-(3-methyl-thiophen-2-yl)-methanone

[0825]

[0826] The title compound was prepared in a manner like that describedfor Example C(1).1-tert-Butoxycarbonyl-4-(4-isothiocyanato-phenyl)-piperazine (fromExample C(101)) and 2-bromoacetyl-3-methyl-thiophene (from ExampleC(19)) gave, after recrystallization with EtOAc/hexane, 387 mg (52%yield) of a yellow solid, mp 175-176° C.

[0827]¹H NMR (CDCl₃): δ 7.00-6.85 (4H, m), 3.62 (4H, dd, J=5.3, 5.0 Hz),3.18 (4H, dd, J=5.3, 5.0 Hz), 2.48 (3H, s), 1.42 (9H, s).

[0828] IR (KBr): 3260, 2978, 1725, 1684, 1601, 1531, 1419, 1231 cm⁻¹.

[0829] Anal. Calcd. for C₂₄H₂₉N₅O₃S₂: C, 57.68; H, 5.85; N, 14.02; S,12.83. Found: C, 57.74; H, 5.82; Cl, N, 13.95; S, 12.95.

Example C(105)4-Amino-2-[4-(1-tert-butoxycarbonyl-piperazine-4-yl)-phenylamino]-thiazol-5-yl-(2,6-dichlorophenyl)-methanone

[0830]

[0831] The title compound was prepared in a manner like that describedfor Example C(1).1-tert-Butoxycarbonyl-4-(4-isothiocyanato-phenyl)-piperazine (fromExample C(101)) and 2-bromo-2′,6′-dichloro-acetophenone (from ExampleC(52)) afforded a crude product, which was purified via flash columnchromatography with MeOH:CH₂Cl₂ (2.5:97.5) as eluant and azeotroped withhexanes to give a yellow solid in 90% yield, mp 165-167° C.

[0832]¹H NMR (CDCl₃): δ 7.22 (2H, d, J=9.0 Hz), 6.92 (2H, d, J=9.0 Hz),3.60 (4H, m), 3.18 (4H, m), 1.42 (9H, s).

[0833] IR (KBr): 3401, 3271, 2966, 1689, 1607, 1542, 1460, 1225 cm⁻¹.

[0834] HRFABMS: Calcd. for C₂₅H₂₈N₅O₃ClS (MH⁺): 548.1290. Found:548.1270.

[0835] Anal. Calcd. for C₂₅H₂₇N₅O₃Cl₂S.0.1C₆H₁₄: C, 55.23; H, 5.07; N,12.58; Cl, 12.74; S, 5.76. Found: C, 55.34; H, 5.28; N, 12.29; Cl,12.48; S. 5.58.

Example C(106)(3-Acetamido-2,6-dichloro-phenyl)-[4-amino-2-(4-tert-butoxycarbonyl-piperazin-1-yl)-amino-thiazol-5-yl]-methanone

[0836]

[0837] The title compound was prepared in a manner like that describedfor Example C(1).1-tert-Butoxycarbonyl-4-(4-isothiocyanato-phenyl)-piperazine (fromExample C(101)) and N-(3-bromoacetyl-2,4-dichloro-phenyl)-acetamide(from Example C(82)) provided a pale yellow solid in 57% yield, mp248-250° C.

[0838]¹H NMR (CDCl₃): δ 7.20 (2H, d, J=9.0 Hz), 6.92 (2H, d, J=9.0 Hz),3.54-3.66 (4H, m), 3.12-3.22 (4H, m), 2.28 (3H, s), 1.42 (9H, s).

[0839] IR (KBr): 3377, 3271, 3177, 2978, 1672, 1548, 1437, 1290, 1231cm⁻¹.

[0840] HRFABMS: Calcd. for C₂₇H₃₁Cl₂N₆O₄S (MH⁺): 605.1505. Found:605.1528.

[0841] Anal. Calcd. for C₂₇H₃₀Cl₂N₆O₄S.1.3H₂O: C, 51.56; H, 5.22; N,13.36; Cl, 11.27; S, 5.10. Found: C, 51.50; H, 5.18; Cl, 11.15; N,13.19; S, 4.99.

Example C(107)4-[4-Amino-5-(2,4,6-trichloro-benzoyl)-thiazol-2-yl-amino]-benzenesulfonamide

[0842]

[0843] 2,4,6-Trichloroacetophenone, which has the structural formula

[0844] was first prepared as follows. Adapted from a procedure byReynolds et al., Org. Syn. Coll., vol. IV (1963), pp. 708-710. To Mgturnings (283 mg, 11.3 mmol) and EtOH (0.25 mL) was added CCl₄ (11 μL).The ensuing reaction subsided, before a solution of diethyl malonate(1.71 mL, 11.33 mmol) in EtOH (0.91 mL) was added at a rate to sustainreaction. After 30 min, the mixture was refluxed to consume Mg for onehour, then allowed to cool. The solid mass was suspended in ether (25mL) and a solution of 2,4,6-trichlorobenzoyl chloride (2.50 g, 10.3 mL)in ether (5 mL) was added cautiously. After 3 days, a solution of H₂SO₄(0.6 mL) in water (10 mL) was carefully added to dissolve any solids,and extracted with ether (2×10 mL). The extracts were dried over MgSO4and evaporated to a cloudy oil, which was placed in HOAc (3 mL), H₂O (2mL) and H₂SO₄ (0.33 mL), and heated to reflux. After 7.5 hours, themixture was allowed to cool overnight. The mixture was made alkalinewith 1N NaOH (35 mL) and extracted with ether (3×10 mL). The combinedethereal layers were dried with MgSO₄ and evaporated to give 1.80 g(78%) of a white solid that was used without further purification(previously described in Baker et al., J. Chem. Soc. (1941), pp.796-802).

[0845] 2-Bromo-2′,4′,6′-trichloroacetophenone, which has the structuralformula

[0846] was prepared in a manner analogous to 2-bromo-2′iodo-acetophenonefor Example C(12). Crude 2′,4′,6′-trichloroacetophenone afforded 1.27 g(94%) of gold crystals that were used without further purification(previously described in Baker et al., J. Chem. Soc. (1941), pp.796-802).

[0847]¹H NMR: δ 7.42 (2H, s), 4.42 (s, 2H).

[0848] The title compound was prepared essentially as described forExample C(1), except that excess potassium t-butoxide (2.2 equivalents)was employed. 4-Isothiocyanato-benzenesulfonamide and2-bromo-2′,4′,6′-trichloroacetophenone provided a dark brown gum, whichwas purified via column chromatography with 10% MeOH/CHCl₃ andprecipitated from MeOH/CHCl₃ to obtain 96 mg (21%) of an amorphous, paleyellow solid.

[0849]¹H NMR (CD₃OD): δ 7.87 (4H, dd, J=14.6, 9.0 Hz), 7.60 (2H, s).

[0850] IR (KBr): 3312, 1593, 1545, 1459, 1421, 1161 cm⁻¹.

[0851] ESIMS (MH⁺): 477/479/481. (M⁻): 475/477/479.

[0852] Anal. Calcd for C₁₆H₁₁Cl₃N₄O₃S₂: C, 40.22; H, 2.32; N, 11.73; Cl,22.26; S, 13.42. Found: C, 40.12; H, 2.34; N, 11.56; Cl, 22.41; S,13.43.

Example C(108)4-[4-Amino-5-(2,6-difluoro-benzoyl)-thiazol-2-ylamino]-N-methyl-benzenesulfonamide

[0853]

[0854] 4-Amino-N-methyl-benzenesulfonamide, which has the structuralformula

[0855] was first made as follows. N-Methyl-4-nitro-benzenesulfonamide(2.58 g, 11.9 mmol; Khanna et al., J. Med. Chem., vol. 40 (1997), pp.1619-1633) and 10% Pd/C (250 mg) in MeOH (60 mL) was stirred underhydrogen atmosphere for 2 hours and filtered. The filtrate wasconcentrated in vacuo to provide 2.17 g (98% yield) of colorlesscrystalline flakes, which by ¹H NMR matched that reported in theliterature (Khanna et al., J. Med. Chem., vol. 40 (1997), pp. 1619-1633)and was used without further purification.

[0856] 4-Isothiocyanato-N-methyl-benzenesulfonamide, which has thestructural formula

[0857] was prepared in a manner analogous to 4-isothiocyanato-benzamideof Example C(102). 4-Amino-N-methyl-benzenesulfonamide (2.17 g, 11.7mmol) gave 2.10 g (79% yield) of white fluffy powder, which was usedwithout further purification.

[0858]¹H NMR (DMSO-d₆): δ 7.83 (2H, d, J=8.4 Hz), 7.65 (2H, d, J=8.4Hz), 7.61 (1H, q, J=4.9 Hz), 2.43 (3H, d, J=4.9 Hz).

[0859] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Isothiocyanato-N-methyl-benzenesulfonamide and2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided acrude product, which was extracted with 10% i-PrOH/CHCl₃ and purifiedvia column chromatography with 5% MeOH/CHCl₃ to afford an amorphousyellow powder in 41% yield, that decomposed above 200° C.

[0860]¹H NMR (DMSO-d₆): δ 11.23 (1H, s), 8.33 (2H, bs), 7.81 (2H, d,J=8.5 Hz), 7.54 (2H, d, J=8.5 Hz), 7.63-7.41 (1H, m), 7.39 (1H, q, J=5.0Hz), 7.23 (2H, t, J=7.1 Hz), 2.41 (3H, d, J=5.0 Hz).

[0861] HRFABMS (MH⁺): Calcd.: 425.0554. Found: 425.0566

[0862] Anal. Calcd. for C₁₇H₁₄N₄O₃S₂F₂.0.5CH₃OH: C, 47.72; H, 3.66; N,12.72; S, 14.56. Found C, 47.56; H, 3.52; N, 12.72; S, 14.77.

Example C(109)4-[4-Amino-5-(2,6-difluoro-benzoyl)-thiazol-2-ylamino]-N,N-dimethyl-benzenesulfonamide

[0863]

[0864] 4-Amino-N,N-dimethyl-benzenesulfonamide, which has the structuralformula

[0865] was next prepared as follows. CrudeN,N-dimethyl-4-nitro-benzenesulfonamide (3.89 g, 16.9 mmol; Khanna etal., J. Med. Chem., vol. 40 (1997), pp. 1619-1633), 10% Pd/C (800 mg),MeOH (80 mL), and THF (200 mL) were stirred under hydrogen for 6 hoursand filtered. The filtrate was concentrated in vacuo to furnish 3.68 gof yellow solid, which was identical by ¹H NMR spectrum to previousdescription by Khanna et al., J. Med. Chem., vol. 40 (1997), pp.1619-1633 and was used without further purification.

[0866] 4-Isothiocyanato-N,N-dimethyl-benzenesulfonamide, which has thestructural formula

[0867] was next made as follows. To a solution of4-amino-N,N-dimethyl-benzenesulfonamide (2.0 g, 10 mmol) in acetone (50mL) at 5-10° C. were added simultaneously a solution of thiophosgene(0.91 mL, 12 mmol) in acetone (20 mL) and 25% aq Na₂CO₃ (10 mL). After 5min at 5-8° C., the mixture was allowed to warm and was stirred atambient temperature for a half hour. The solvent was evaporated andwater (70 mL) was added. The resultant light-yellow precipitate wasfiltered off, washed with water, and dried under vacuum to afford 2.35 g(97% yield) of white powder, which was used without furtherpurification.

[0868]¹H NMR (DMSO-d₆): δ 7.82 (2H, d, J=8.4 Hz), 7.69 (2H, d, J=8.4Hz), 2.63 (6H, s).

[0869] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Isothiocyanato-N,N-dimethyl-benzenesulfonamideand 2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided acrude brown solid that recrystallized from EtOH to give light-browncrystals in 52% yield, mp 240-242° C.

[0870]¹H NMR (DMSO-d₆): δ 11.24 (1H, s), 8.14 (2H, bs), 7.84 (2H, d,J=8.8 Hz), 7.72 (2H, d, J=8.8 Hz), 7.62-7.49 (1H, m), 7.23 (1H, d, J=7.9Hz), 7.20 (1H, d, J=8.0 Hz), 2.59 (6H, s).

[0871] Anal. Calcd. for C₁₈H₁₆N₄O₃S₂F₂: C, 49.31; H, 3.68; N, 12.78; S,14.63. Found: C, 49.29; H, 3.71; N, 12.68; S, 14.50.

Example C(110)(4-Amino-2-{4-[(2-dimethylamino-ethyl)-methyl-amino]-phenylamino}-thiazol-5-yl)-(2,6-difluoro-phenyl)-methanone

[0872]

[0873] N-(4-Nitrophenyl)-N,N′,N′-trimethyl-ethane-1,2-diamine, which hasthe strutural formula

[0874] was first prepared in a manner analogous to tert-butyl[methyl-(4-nitro-phenyl)-amino]-acetate for Example C(103).4-Fluoronitrobenzene and N,N,N′-trimethyl-ethylendiamine gave a brownoil in 87% crude yield, which was used without any further purification.

[0875]¹H NMR (CDCl₃): δ 8.14 (2H, d, J=9.6 Hz), 6.64 (2H, d, J=9.3 Hz),3.58 (2H, t, J=7.5 Hz), 3.12 (3H, s) 2.52 (2H, t, J=7.5 Hz), 2.32 (6H,s).

[0876] N-(4-Aminophenyl)-N,N′,N′-ethane-1,2-diamine, which has thestructural formula

[0877] was prepared in a manner analogous to4-(4-methyl-piperazin-1-yl)-aniline for Example C(70).N-(4-nitrophenyl)-N,N,N′-trimethyl-ethane-1,2-diamine furnished areddish-brown oil in 92% crude yield which was used without furtherpurification.

[0878]¹H NMR (CDCl₃): δ 6.62 (4H, s), 3.30 (2H, dd, J=7.6, 7.4 Hz), 2.85(3H, s), 2.47 (2H, dd, J=7.7, 7.2 Hz), 2.32 (6H, s).

[0879] N-(4-Isothiocyanato-phenyl)-N,N′,N′-trimethyl-ethane-1,2-diamine,which has the structural formula

[0880] was prepared in a manner analogous to1-(4-isothiocyanato-phenyl)-1H-imidazole for Example C(41).N-(4-Aminophenyl)-N,N′,N′-ethane-1,2-diamine provided a brown oil in 75%crude yield, which was used without further purification.

[0881]¹H NMR (CDCl₃): δ 7.13 (2H, d, J=8.8 Hz), 7.01 (2H, d, J=8.2 Hz),3.99 (2H, dd, J=7.6, 7.1 Hz), 3.15 (1H, bs), 3.02 (3H, s), 2.80 (6H, s).

[0882] The title compound was prepared in a manner like that describedfor Example C(1).N-(4-Isothiocyanato-phenyl)-N,N′,N′-trimethyl-ethane-1,2-diamine and2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) afforded acrude product, which was purified via flash column chromatography with astepwise gradient of MeOH:CH₂Cl₂ (2.5:97.5-10:90) to provide a yellowsolid in 55% yield, mp 96-98° C.

[0883]¹H NMR (DMSO-d₆): δ 7.42-7.55 (1H, m), 7.10-7.24 (4H, m), 6.64(2H, d, J=9.0 Hz), 2.90 (3H, s), 2.38 (2H, dd, J=7.2, 6.5 Hz), 2.18 (6H,s).

[0884] IR (KBr): 3394, 3180, 2948, 2828, 1620, 1546, 1523, 1466 cm⁻¹.

[0885] HRFABMS: Calcd. for C₂₁H₂₄F₂N₅OS (MH⁺): 432.1670. Found:432.1658.

[0886] Anal. Calcd. for C₂₁H₂₃F₂N₅OS.0.4H₂O: C, 57.49; H, 5.47; N,15.96; S, 7.31. Found: C, 57.36; H, 5.45; N, 15.77; S, 7.27.

Example C(111)2-[4-(1-Acetyl-piperazin-4-yl)-phenylamino]-4-amino-thiazol-5-yl-(2,6-difluorophenyl)-methanone

[0887]

[0888] The title compound was prepared in a manner like that describedfor Example C(1). 1-Acetyl-4-(4-isothiocyanato-phenyl)-piperazine (fromExample C(97)) and 2-bromo-2′,6′-difluoro-acetophenone (from ExampleC(79)) provided 320 mg (66% yield) of a cream-colored solid, mp 298° C.

[0889]¹H NMR (DMSO-d₆): δ 7.44-7.58 (1H, m), 7.36 (2H, bd, J=7.2 Hz),7.18 (2H, dd, J=8.1, 7.5 Hz), 6.95 (2H, d, J=9.0 Hz), 3.58 (4H, bs),3.00-3.20 (4H, m), 2.05 (3H, s)

[0890] IR (KBr): 3389, 3154, 1607, 1601, 1542, 1419, 1231 cm⁻¹.

[0891] HRFABMS: Calcd. for C₂₂H₂₁F₂N₅ONa (M+Na⁺) 480.1282. Found:480.1266.

[0892] Anal. Calcd. for C₂₂H₂₁N₅O₂F₂S.0.3H₂O: C, 57.08; H, 4.70; N,15.13; S, 6.93. Found: C, 56.95; H, 4.74; N, 15.16; S, 6.82

Example C(112)2-[4-(1-Acetyl-piperazin-4-yl)-phenylamino]-4-amino-thiazol-5-yl-(2,5-dimethyl-thiophen-3-yl)-methanone

[0893]

[0894] The title compound was prepared in a manner like that describedfor Example C(1). 1-Acetyl-4-(4-isothiocyanato-phenyl)-piperazine (fromC(97)) and 3-bromoacetyl-2,5-dimethyl-thiophene (from Example C(50))provided 200 mg (53% yield) of a pale cream-colored solid, mp 282-283°C.

[0895]¹H NMR (DMSO-d₆): δ 7.42 (2H, d, J=9.0 Hz), 6.98 (2H, d, J=9.0Hz), 6.82 (1H, s), 3.60 (4H, bs), 3.02-3.20 (4H, m), 2.46 (3H, s), 2.38(3H, s), 2.05 (3H, s).

[0896] IR (KBr): 3401, 3166, 1637, 1601, 1542, 1425, 1231 cm⁻¹.

[0897] HRFABMS: Calcd. for C₂₂H₂₆N₅O₂S₂ (MH⁺): 456.1528. Found:456.1510.

[0898] Anal. Calcd. for C₂₂H₂₅N₅O₂S₂: C, 57.87; H, 5.74; N, 15.34; S,14.05. Found: C, 57.85; H, 5.53; N, 15.23; S, 14.20.

Example C(113)4-[4-Amino-5-(2-fluoro-6-trifluoromethyl-benzoyl)-thiazol-2-ylamino]-benzamide

[0899]

[0900] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Isothiocyanato-benzamide (from Example C(102))and 2-bromo-2′-fluoro-6′-trifluoromethyl-acetophenone (from ExampleC(99)) provided a crude product, which was purified via columnchromatography with a stepwise gradient of 8-10% EtOH/CHCl₃ to afford anamorphous yellow solid in 14% yield that decomposed above 145° C.

[0901]¹H NMR (DMSO-d₆): δ 8.30 (1H, bs), 8.10 (1H, bs), 7.94-7.82 (3H,m), 7.74-7.62 (5H, m), 7.30 (1H, s).

[0902] HRFABMS (MH⁺): Calcd.: 425.0695. Found: 425.0709.

[0903] Anal. Calcd. for C₁₈H₁₂N₄O₂SF₄.0.9EtOH: C, 51.05; H,3.76; N,12.03; S, 6.88. Found: C, 51.14; H, 3.78; N, 12.36; S, 6.79.

Example C(114)4-[4-Amino-5-(3-methyl-thiophene-2-carbonyl)-thiazol-2-ylamino]-N-methyl-benzenesulfonamide

[0904]

[0905] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-Isothiocyanato-N-methyl-benzenesulfonamide (fromExample C(108)) and 2-bromoacetyl-3-methyl-thiophene (from ExampleC(19)) provided a yellow solid in 57% yield, mp 197.0-199.5° C.

[0906]¹H NMR (DMSO-d₆): δ 11.19 (1H, s), 8.24 (2H, bs), 7.86 (2H, d,J=8.7 Hz), 7.75 (2H, d, J=8.7 Hz), 7.65 (1H, d, J=5.0 Hz), 7.36 (1H, q,J=6.1 Hz), 7.03 (1H, d, J=5.0 Hz), 2.42 (3H, S), 2.41 (3H, d, J=6.1 Hz).

[0907] HRFABMS (MH⁺): Calcd.: 409.0463. Found: 409.0474.

[0908] Anal. Calcd. for C₁₆H₁₆N₄O₃S₃.0.4H₂O: C, 46.23; H, 4.07; N,13.48; S, 23.14. Found: C, 46.28; H, 3.98; N, 13.38; S, 23.08.

Example C(115)4-[4-Amino-5-(2,4,6-trifluoro-benzoyl)-thiazol-2-yl-amino]-benzenesulfonamide

[0909]

[0910] 2-Chloro-2′,4′,6′-trifluoroacetophenone, which has the structuralformula

[0911] was first prepared as follows. To a mechanically stirred solutionof 1,3,5-trifluorobenzene (5.17 mL, 50.0 mmol) in dichloroethane (12.5mL) was added gradually AlC₃ (13.4 g, 115 mmol) over 15 min. time periodwith caution. Violent bumping and HCl gas evolution was observed. Themixture was carefully heated to reflux, and chloroacetyl chloride (6.20g, 4.37 mL, 55.0 mmol) was added dropwise over 45 min. time period.After 6 hours at reflux, the mixture was allowed to cool over 12 hours,then carefully poured onto an ice/water slush (˜200 mL) and extractedwith ether (3×50 mL). The combined ethereal layers were washed with 10%aq. HCl (2×30 mL), 1N aq. NaOH (3×30 mL), and brine (25 mL), dried overMgSO4 and evaporated to give 5.28 g (51%) of a yellow solid that wasused without further purification. (An analytical sample crystallizedfrom ether/hexane to give yellow microcrystals, mp 43-45° C.)

[0912]¹H NMR (CDCl₃): δ 6.81 (2H, t, J=8.4 Hz), 4.54 (2H, s).

[0913] IR(KBr): 1721, 1637, 1616, 1447, 1201, 1128, 1045 cm⁻¹.

[0914] Anal. Calcd. for C₈H₄ClF₃O: C, 46.07; H, 1.93; Cl, 17.00. Found:C, 45.92; H, 1.95; Cl, 16.97.

[0915] The title compound was prepared essentially as described forExample C(1), except that excess potassium t-butoxide (2.2 equivalents)was employed. 4-Isothiocyanato-benzenesulfonamide and2-chloro-2′,4′,6′-trifluoroacetophenone gave a red-brown solid, whichwas purified via column chromatography with 5% MeOH/CH₂Cl₂ as eluant.Precipitation with trace hexane in MeOH/CH₂Cl₂ gave 70 mg (33%) ofyellow amorphous powder that decomposed above 148° C.

[0916]¹H NMR (CD₃OD): δ 7.91 (1H, s), 7.86 (4H, dd, J=14.9, 6.9 Hz),6.99 (2H, dd, J=9.0, 7.5 Hz).

[0917] IR(KBr): 3278, 1602, 1549, 1425, 1155 cm⁻¹.

[0918] HRFABMS. Calcd for C₁₆H₁₂F₃N₄O₃S₂ (MH⁺): 429.0303. Found:429.0315.

[0919] Anal. Calcd for C₁₆H₁₁F₃N₄O₃S₂.1.1H₂O: C, 42.87; H, 2.97; N,12.50; S, 14.31. Found: C, 42.98; H, 2.73; N, 12.12; S, 14.48.

Example C(116){4-Amino-2-[4-(4-methyl-piperazine-1-sulfonyl)-phenylamino]-thiazol-5-yl}-(2,6-difluoro-phenyl)-methanone

[0920]

[0921] 1-Methyl-4-(4-nitro-benzenesulfonyl)-piperazine, which has thestructural formula

[0922] was prepared in a manner analogous to that used forN-methyl-4-nitro-benzenesulfonamide for Example C(108) (Khanna et al.,J. Med. Chem., vol. 40 (1997), pp. 1619-1633). 4-Nitrobenzenesulfonylchloride and 1-methylpiperazine gave 5.1 g (88% yield) of yellow solid,which was used without further purification.

[0923] 4-(4-Methyl-piperazine-1-sulfonyl)-aniline, which has thestructural formula

[0924] was prepared in a manner analogous to that used forN-methyl-4-amino-benzenesulfonamide for Example C(108).1-Methyl-4-(4-nitro-benzenesulfonyl)-piperazine provided a gray solid in99% yield, which was used in the next step without further purification.

[0925]¹H NMR (DMSO-d₆): δ 7.37 (2H, d, J=8.8 Hz), 6.67 (2H, d, J=8.8Hz), 6.16 (2H, bs), 3.30 (4H, bs), 3.03 (4H, bs), 2.58 (3H, s).

[0926] 1-(4-Isothiocyanato-benzenesulfonyl)-4-methyl-piperazine, whichhas the structural formula

[0927] was made in a manner analogous to 4-isothiocyanato-benzamide forExample C(102). 4-(4-Methyl-piperazine-1-sulfonyl)-aniline provided 1.1g (94% yield) of white crystals which were used without furtherpurification.

[0928]¹H NMR (CDCl₃): δ 7.74 (2H, d, J=8.6 Hz), 7.35 (2H, d, J=8.6 Hz),3.27 (4H, bs), 2.77 (4H, bs), 2.47 (3H, s).

[0929] The title compound was prepared in a manner analogous to thatused in Example C(1).1-(4-Isothiocyanato-benzenesulfonyl)-4-methyl-piperazine and2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided ayellow solid in 69% yield, mp 172-174° C.

[0930]¹H NMR (DMSO-d₆): δ 11.23 (1H, bs), 8.21 (2H, bs), 7.84 (2H, d,J=8.8 Hz), 7.69 (2H, d, J=8.8 Hz), 7.62-7.49 (11H, m), 7.22 (1H, d,J=7.8 Hz), 7.19 (11H, d, J=8.1 Hz), 2.87 (4H, t, J=4.5 Hz), 2.35 (4H, t,J=4.5 Hz), 2.13 (3H, s).

[0931] HRFABMS (MH⁺): Calcd.: 494.1132. Found: 494.1120.

[0932] Anal. Calcd. for C₂₁H₂₁N₅O₃S₂F₂.0.1H₂O.0.5 CH₃OH: C, 50.50; H,4.57; N, 13.70; S, 12.54. Found: C, 50.34; H, 4.39; N, 13.51; S, 12.63.

Example C(117)(4-Amino-2-{4-[(2-dimethylamino-ethyl)-methyl-amino]-phenylamino}-thiazol-5-yl)-(3-methyl-thiophen-2-yl)-methanone

[0933]

[0934] The title compound was prepared in a manner like that describedfor Example C(1).N-(4-Isothiocyanato-phenyl)-N,N′,N′-trimethyl-ethane-1,2-diamine (fromExample C(110)) and 2-bromoacetyl-3-methyl-thiophene (from ExampleC(19)) gave, after purification via flash column chromatography withMeOH:CH₂Cl₂ (5:95) as eluant, a yellow foam in 70% yield.

[0935]¹H NMR (DMSO-d₆): 67.22 (1H, d, J=5.0 Hz), 7.16 (2H, d, J=9.0 Hz),6.72 (1H, d, J=5.0 Hz), 6.58 (2H, d, J=9.0 Hz), 3.44 (2H, dd, J=7.7, 7.4Hz), 3.00 (3H, s), 2.42 (3H, s), 2.3 (6H, s).

[0936] IR (KBr): 3377, 3269, 2937, 2821, 1609, 1543, 1518, 1423 cm⁻¹.

[0937] HRFABMS: Calcd. for C₂₀H₂₆Cl₂N₅OS₂ (MH⁺): 416.1579. Found:416.1594.

[0938] Anal. Calcd. for C₂₀H₂₅Cl₂N₅OS₂.1H₂O: C, 55.40; H, 6.28; N,16.15; S, 14.71. Found: C, 55.43; H, 5.94; N, 16.37; S, 14.57.

Example C(118)4-{4-[4-Amino-5-(2,6-difluoro-benzoyl)-thiazol-2-ylamino]-phenyl}-1-methyl-piperazin-2-one

[0939]

[0940] 4-(4-Nitro-phenyl)-piperazin-2-one, which has the structuralformula

[0941] was first prepared in a manner analogous to tert-butyl[methyl-(4-nitro-phenyl)-amino]-acetate for Example C(103).Piperazin-2-one (Aspinall et al., J. Amer. Chem. Soc., vol. 62 (1940),pp. 1202-1204) and 4-fluoronitrobenzene furnished a yellow solid in 63%yield, which was used without any further purification.

[0942]¹H NMR (CDCl₃): δ 8.10 (2H, d, J=8.8 Hz), 6.80 (2H, d, J=9.2 Hz),6.38 (1H, bs), 4.10 (2H, s), 3.74-2.52 (4H, m).

[0943] 1-Methyl-4-(4-nitro-phenyl)-piperazin-2-one, which has thestructural formula

[0944] was next prepared as follows. To a suspension of4-(4-nitro-phenyl)-piperazin-2-one (500 mg, 2.26 mmol) in THF (5 mL) wasadded NaH (60 mg, 2.5 mmol). The mixture was cooled to 0° C.,iodomethane (162 uL, 2.59 mmol) was added, and then the mixture wasallowed to warm to ambient temperature. After 12 hours, the solvent wasremoved in vacuo to give a yellow gum, which was treated with H₂O. Theresultant yellow precipitate was filtered off, washed with H₂O, anddried under high vacuum for several hours to afford 420 mg (79% yield).

[0945]¹H NMR (CDCl₃): δ 8.18 (2H, d, J=9.4 Hz), 6.78 (2H, d, J=9.4 Hz),4.08 (2H, s), 3.68 (2H, dd, J=4.7, 3.6 Hz), 3.54 (2H, dd, J=4.9, 3.7Hz), 3.02 (3H, s).

[0946] 4-(4-Amino-phenyl)-1-methyl-piperazin-2-one, which has thestructural formula

[0947] was prepared in a manner analogous to4-(4-methyl-piperazin-1-yl)-aniline for Example C(70).1-Methyl-4-(4-nitro-phenyl)-piperazin-2-one provided a brown gum, whichwas used without any further purification.

[0948]¹H NMR (CDCl₃): δ 6.78 (2H, d, J=9.0 Hz), 6.60 (2H, d, J=9.0 Hz),3.76 (2H, s), 3.44 (2H, dd, J=5.8, 4.9 Hz), 3.20 (2H, dd, J=4.9, 4.0Hz), 3.02 (3H, s).

[0949] 4-(4-Isothiocyanato-phenyl)-1-methyl-piperazin-2-one, which hasthe structural formula

[0950] was prepared in a manner analogous to1-(4-isothiocyanato-phenyl)-1H-imidazole for Example C(41).4-(4-Amino-phenyl)-1-methyl-piperazin-2-one gave a cream-colored powderin 85% yield, which was used without further purification.

[0951]¹H NMR (CDCl₃): δ 7.18 (2H, d, J=9.0 Hz), 6.80 (2H, d, J=9.0 Hz),3.90 (2H, s), 3.50 (4H, bs), 3.70 (3H, s).

[0952] The title compound was prepared in a manner like that describedfor Example C(1). 2-bromo-2′,6′-difluoro-acetophenone (from ExampleC(79)) and 4-(4-isothiocyanato-phenyl)-1-methyl-piperazin-2-one provideda yellow solid in 77% yield, mp>300° C.

[0953]¹H NMR (DMSO-d₆): δ 7.60-7.70 (1H, m), 7.48 (2H, bd, J=8.3 Hz),7.31 (2H, t, J=7.9 Hz), 7.09 (2H, d, J=9.0 Hz), 3.88 (2H, s), 3.58 (4H,bd, J=4.4 Hz), 3.02 (3H, s).

[0954] Anal. Calcd. for C₂₁H₁₉F₂N₅O₂S: C, 56.88; H, 4.32; N, 15.79; S,7.23. Found: C, 56.81; H, 4.42; N, 15.83; S, 7.31.

Example C(119)[4-Amino-2-(4-thiomorpholin-4-yl-phenylamino)-thiazol-5-yl]-(2,6-difluoro-phenyl)-methanone

[0955]

[0956] 4-Thiomorpholin-4-yl-aniline, which has the structural formula

[0957] was first prepared as follows. 4-(4-Nitro-phenyl)-thiomorpholine(1.50 g, 6.70 mmol; Beach et al., J. Chem. Soc. Perkin Trans. 2 (1984),pp. 217-221) and 10% Pd/C (200 mg of wet DeGussa type, 50% by wt.) wasstirred in ethyl acetate (20 mL) and MeOH (20 mL) under hydrogenovernight and filtered. The filtrate was concentrated in vacuo to give1.28 g (98% yield) of white crystalline flakes, which were used withoutfurther purification.

[0958] 4-(4-Isothiocyanato-phenyl)-thiomorpholine, which has thestructural formula

[0959] was prepared in a manner analogous to4-isothiocyanato-N,N-dimethyl-benzenesulfonamide for Example C(109).4-Thiomorpholin-4-yl-aniline provided a yellow powder in 83% yield.

[0960]¹H NMR (CDCl₃): δ 7.13 (2H, d, J=9.1 Hz), 6.79 (2H, d, J=9.1 Hz),3.59 (4H, ddd, J=5.2, 5.0, 2.6 Hz), 2.72 (4H, ddd, J=5.2, 5.0, 2.6 Hz).

[0961] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-(4-Isothiocyanato-phenyl)-thiomorpholine and2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided ayellow powder in 51% yield, mp 128-130° C.

[0962]¹H NMR (DMSO-d₆): δ 10.64 (1H, s), 8.12 (2H, bs), 7.56-7.44 (1H,m), 7.30 (2H, d, J=9.0 Hz), 7.18 (1H, d, J=7.7 Hz), 7.15 (1H, d, J=8.1Hz), 6.91 (2H, d, J=9.0 Hz), 3.47 (2H, dd, J=5.1, 5.0 Hz), 2.65 (2H, dd,J=5.1, 5.0 Hz).

[0963] HRFABMS (MH⁺): Calcd.: 433.0968. Found: 433.0980.

[0964] Anal. Calcd. for C₂₀H₁₈N₄OS₂F₂.0.2H₂O: C, 55.08; H, 4.25; N,12.85; S, 14.71. Found: C, 55.02; H, 4.14; N, 12.72; S, 14.53.

Example C(120)4-{4-[4-Amino-5-(2,6-difluoro-benzoyl)-thiazol-2-ylamino]-phenyl}-piperazin-2-one

[0965]

[0966] 4-(4-Amino-phenyl)-piperazin-2-one, which has the structuralformula

[0967] was prepared in a manner analogous to4-(4-methyl-piperazin-1-yl)-aniline for Example C(70).4-(4-Nitro-phenyl)-piperazin-2-one (from Example C(115)) gave a palebrown oil in 100% crude yield, which was used without any furtherpurification.

[0968]¹H NMR (CD₃OD): δ 7.02 (2H, d, J=8.7 Hz), 6.91 (2H, d, J=8.8 Hz),3.81 (2H, s), 3.59 (2H, dd, J=5.9, 4.8 Hz), 3.46 (2H, dd, J=5.9, 4.8Hz).

[0969] 4-(4-Isothiocyanato-phenyl)-piperazin-2-one, which has thestructural formula

[0970] was prepared in a manner analogous to1-(4-isothiocyanato-phenyl)-1H-imidazole for Example C(41).4-(4-Amino-phenyl)-piperazin-2-one provided a cream-colored solid, whichwas used without further purification.

[0971]¹H NMR (CDCl₃): δ 9.00 (1H, bs), 8.20 (2H, d, J=9.0 Hz), 7.80 (2H,d, J=9.0 Hz), 4.50 (2H, s), 4.00-4.30 (4H, m).

[0972] The title compound was prepared in a manner like that describedfor Example C(1). 2-bromo-2′,6′-difluoro-acetophenone (from ExampleC(79)) and 4-(4-isothiocyanato-phenyl)-piperazin-2-one provided a yellowsolid in 56% yield, mp 280-282° C.

[0973]¹H NMR (DMSO-d₆): δ 9.12 (3H, bs), 8.32-8.44 (1H, m), 8.18 (2H,bd, J=6.9 Hz), 8.05 (2H, t, J=8.2 Hz), 7.78 (2H, d, J=9.0 Hz), 4.52 (2H,s).

[0974] HRFABMS: Calcd. for C₂₀H₁₈F₂N₅O₂S (MH⁺): 430.1149. Found:430.1138.

[0975] Anal. Calcd. for C₂₀H₁₇F₂N₅O₂S.0.3H₂O: C, 55.24; H, 4.08; N,16.11; S, 7.37. Found: C, 55.24; H, 4.10; N, 15.87; S, 7.34.

Example C(121){4-Amino-2-[4-(4-cyclopropylmethyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(2,6-difluoro-phenyl)-methanone

[0976]

[0977] 1-Cyclopropylmethyl-4-(4-nitro-phenyl)-piperazine, which has thestructural formula

[0978] was first prepared as follows. To a suspension of1-(4-nitro-phenyl)-piperazine (2.50 g, 12.1 mmol) in DMF (10 mL) wasadded anhydrous Na₂CO₃ (639 mg, 6.03 mmol) and bromomethylcyclopropane(585 μL, 6.03 mmol). The mixture was heated at 100° C. overnight, thenallowed to cool and diluted with H₂O (30 mL). The separated aqueouslayer was extracted with CHCl₃ (3×50 mL). The combined organic layerswere dried over Na₂SO₄, filtered, and concentrated under reducedpressure to give an orange-brown solid, which was purified via flashcolumn chromatography with 2.5% MeOH/CH₂Cl₂ as eluant to give 2.65 g(84% yield) of a yellow solid. This material was used without anyfurther purification.

[0979]¹H NMR (CDCl₃): δ 8.10 (2H, d, J=10.7 Hz), 7.11 (2H, d, J=9.5 Hz),3.45 (4H, dd, J=5.3, 5.1 Hz), 2.65 (4H, dd, J=5.3, 5.1 Hz), 2.29 (2H, d,J=6.6 Hz), 0.84-0.98 (1H, m), 0.50-0.58 (2H, m), 0.10-0.15 (2H, m).

[0980] 4-(4-Cyclopropylmethyl-piperazin-1-yl)-aniline, which has thestructural formula

[0981] was prepared in a manner analogous to4-(4-methyl-piperazin-1-yl)-aniline for Example C(70).1-Cyclopropylmethyl-4-(4-nitro-phenyl)-piperazine furnished a red solidin 99% crude yield, which was used without further purification.

[0982]¹H NMR (CDCl₃): δ 6.85 (2H, d, J=9.9 Hz), 6.62 (2H, d, J=8.8 Hz),3.42 (2H, bs), 3.10 (4H, dd, J=5.1, 4.8 Hz), 2.69 (4H, dd, J=5.1, 4.9Hz), 2.30 (2H, d, J=6.5 Hz), 0.90-0.98 (1H, m), 0.50-0.56 (2H, m),0.10-0.15 (2H, m).

[0983] 1-Cyclopropylmethyl-4-(4-isothiocyanato-phenyl)-piperazine, whichhas the structural formula

[0984] was prepared in a manner analogous to1-(4-isothiocyanato-phenyl)-1H-imidazole for Example C(41).4-(4-Cyclopropylmethyl-piperazin-1-yl)-aniline gave a dark-brown oil in95% crude yield, which was used without further purification.

[0985]¹H NMR (CDCl₃): δ 6.80 (2H, d, J=9.0 Hz), 6.68 (2H, d, J=9.1 Hz),3.08 (4H, bs), 2.55 (4H, bs), 2.10 (2H, d, J=6.2 Hz), 0.65-0.80 (1H, m),0.42 (2H, d, J=8.0 Hz), 0.00 (2H, d, J=4.6 Hz).

[0986] The title compound was prepared in a manner like that describedfor Example C(1).1-Cyclopropylmethyl-4-(4-isothiocyanato-phenyl)-piperazine and2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided, aftercrystallization from EtOH, a yellow solid in 17% yield, mp 215-218° C.

[0987]¹H NMR (DMSO-d₆): δ 10.60 (1H, s), 8.04 (2H, bs), 7.46-7.56 (1H,m), 7.18-7.20 (2H, m), 7.08 (2H, dd, J=8.0, 7.7 Hz), 6.82 (2H, d, J=9.1Hz), 2.98-3.03 (4H, m), 2.47 (4H, bs), 2.12 (2H, d, J=6.6 Hz), 0.72-0.78(1H, m), 0.34-0.42 (2H, m), 0.00-0.12 (2H, m).

[0988] IR (KBr): 2917, 1620, 1513, 1428 cm⁻¹.

[0989] HRFABMS: Calcd. for C₂₄H₂₅F₂N₅OSCs (M+Cs⁺): 602.0802. Found:602.0818.

[0990] Anal. Calcd. for C₂₄H₂₅F₂N₅OS.0.5H₂O.0.1EtOH: C, 60.16; H, 5.55;N, 14.49; S, 6.64. Found: C, 59.94; H, 5.24; N, 14.19; S, 6.92.

Example C(122)[4-Amino-2-(4-pyridin-4-yl-phenylamino)-thiazol-5-yl]-(2,6-difluoro-phenyl)-methanone

[0991]

[0992] 4-Pyridin-4-yl-aniline, which has the structural formula

[0993] was first prepared as follows. A mixture of4-(4-nitro-phenyl)-pyridine (600 mg, 3.0 mmol; Wang et al., J. Phys.Chem., vol. 99 (1995), pp. 6876-6888) and 10% Pd/C (100 mg) in EtOH (20mL) was stirred under a hydrogen atmosphere overnight. The catalyst wasfiltered off and the filtrate concentrated in vacuo to provide 510 mg(100% yield) of white solid.

[0994]¹H NMR (CDCl₃): δ 8.59 (2H, dd, J=6.2, 1.6 Hz), 7.51 (2H, d, J=8.6Hz), 7.46 (2H, dd, J=6.2, 1.6 Hz), 6.79 (2H, d, J=8.6 Hz).

[0995] 4-(4-Isothiocyanato-phenyl)-pyridine, which has the structuralformula

[0996] was prepared as follows. To 4-pyridin-4-yl-aniline (200 mg, 1.18mmol) in THF (35 mL) at 0° C. was added in succession Et₃N (0.33 mL, 2.4mmol) and thiophosgene (99 μl, 1.29 mmol) dropwise. After 20 min. at 0°C., then ambient temperature for 10 min., the solvent was evaporated.The residue was suspended in water, filtered, washed with minimal water,and dried under vacuum to give a brown solid, 240 mg (96%), which wasused without further purification.

[0997]¹H NMR (CDCl₃): δ 8.62 (2H, d, J=6.3 Hz), 7.57 (2H, d, J=8.6 Hz),7.45 (2H, d, J=6.3 Hz), 7.27 (2H, d, J=8.6 Hz).

[0998] The title compound was prepared in a manner analogous to thatused in Example C(1). 4-(4-Isothiocyanato-phenyl)-pyridine and2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided, afterrecrystallization from EtOH, a brown powder in 64% yield, mp>300° C.

[0999]¹H NMR (DMSO-d₆): δ 11.08 (1H, s), 8.61 (2H, d, J=6.0 Hz), 8.25(2H, bs), 7.85 (2H, d, J=8.8 Hz), 7.73 (2H, d, J=8.8 Hz), 7.71 (2H, d,J=6.0 Hz), 7.61-7.49 (1H, m), 7.23 (1H, d, J=7.7 Hz), 7.20 (1H, d, J=8.1Hz).

[1000] HRFABMS (MH⁺): Calcd.: 409.0935. Found: 409.0921.

[1001] Anal. Calcd. for C₂₁H₁₄N₄OSF₂.0.4H₂O.0.3EtOH: C, 60.41; H, 3.90;N, 13.05; S, 7.47. Found: C, 60.51; H, 3.65; N, 12.69; S, 7.86.

Example C(123){4-Amino-2-[4-(4-carbamoyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(2,6-difluoro-phenyl)-methanone

[1002]

[1003] 1-Carbamoyl-4-(4-nitro-phenyl)-piperazine, which has thestructural formula

[1004] was first obtained according to a procedure from Cain et al., J.Med. Chem., vol. 20 (1977), pp. 987-996, wherein1-(4-nitrophenyl)piperazine was treated with potassium cyanate toprovide a white solid, 705 mg (99%), which was used without furtherpurification.

[1005]¹H NMR (CDCl₃): δ 6.71 (2H, d, J=8.6 Hz), 6.50 (2H, d, J=8.6 Hz),5.97 (2H, bs), 4.58 (2H, bs), 3.39 (4H, dd, J=5.1, 4.9 Hz), 2.82 (4H,dd, J=5.1, 4.9 Hz).

[1006] 1-(4-Amino-phenyl)-4-carbamoyl-piperazine, which has the formula

[1007] was next prepared as follows. A mixture of4-(4-nitro-phenyl)-piperazine-1-carboxylic acid amide (760 mg, 3.22mmol), 10% Pd/C (120 mg), MeOH (20 mL), and THF (20 mL) was stirredunder hydrogen for 2 hours. The catalyst was filtered off and thefiltrate concentrated in vacuo to provide a white solid, 705 mg (99%),which was used without further purification.

[1008]¹H NMR (CDCl₃): δ 6.71 (2H, d, J=8.6 Hz), 6.50 (2H, d, J=8.6 Hz),5.97 (2H, bs), 4.58 (2H, bs), 3.39 (4H, dd, J=5.1, 4.9 Hz), 2.82 (4H,dd, J=5.1, 4.9 Hz).

[1009] 1-Carbamoyl-4-(4-isothiocyanato-phenyl)-piperazine, which has thestructural formula

[1010] was prepared as follows. To a suspension of1-(4-amino-phenyl)-4-carbamoyl-piperazine (300 mg, 1.36 mmol) in THF (30mL) at −35° C. was successively added triethylamine (0.38 mL, 2.73 mmol)and thiophosgene (104 μl, 1.36 mmol) dropwise. The solvent wasevaporated and the tarry residue diluted with water. The resultant lightbrown solid was filtered off, washed with a small amount of water, anddried under vacuum to afford a brown powder, 337 mg (94% yield), whichwas used without further purification.

[1011]¹H NMR (CDCl₃): δ 7.08 (2H, d, J=9.0 Hz), 6.76 (2H, d, J=9.0 Hz),4.45 (2H, bs), 3.50 (4H, dd, J=5.4, 5.0 Hz), 3.15 (4H, dd, J=5.4, 5.0Hz).

[1012] The title compound was prepared in a manner analogous to thatused in Example C(1). 1-Carbamoyl-4-(4-isothiocyanato-phenyl)-piperazineand 2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided alight-gray powder in 45% yield, mp 278.5-279° C.

[1013]¹H NMR (DMSO-d₆): δ 10.69 (1H, s), 8.16 (2H, bs), 7.63-7.51 (1H,m), 7.38 (2H, d, J=9.0 Hz), 7.25 (1H, d, J=7.8 Hz), 7.21 (1H, d, J=7.9Hz), 7.02 (2H, d, J=9.0 Hz), 6.09 (2H, bs), 3.48 (2H, t, J=4.7 Hz), 3.11(2H, t, J=4.7 Hz).

[1014] HRFABMS (M+Na⁺): Calcd.: 81.1234. Found: 481.1246.

[1015] Anal. Calcd. for C₂₁H₂₀N₆O₂SF₂.0.5H₂O: C, 53.95; H, 4.53; N,17.98; S, 6.86. Found: C, 53.92; H, 4.35; N, 17.64; S, 6.64.

Example C(124){4-Amino-2-[4-(3R,4-dimethyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(2,6-difluoro-phenyl)-methanone

[1016]

[1017] 3R-Methyl-1-(4-nitro-phenyl)-piperazine, which has the structuralformula

[1018] was made first as follows. (R)-(−)-2-Methylpiperazine (186 mg,1.86 mmol), 1-fluoro-4-nitrobenzene (131 mg, 0.93 mmol), Et₃N (0.26 mL,1.86 mmol), and acetonitrile (2 mL) was refluxed overnight and thenconcentrated in vacuo. The residue was suspended in water and theresultant solid was filtered off, washed with minimal water, and driedunder vacuum to provide a bright yellow solid 128 mg (62% yield), whichwas used without further purification.

[1019]¹H NMR (CDCl₃): δ 8.12 (2H, d, J=9.5 Hz), 6.82 (2H, d, J=9.5 Hz),3.80-3.71 (2H, m), 3.18-3.08 (1H, m), 3.04-2.88 (3H, m), 2.58 (1H, dd,J=12.3, 12.3 Hz), 1.16 (3H, d, J=6.3 Hz).

[1020] 1,2R-Dimethyl-4-(4-nitro-phenyl)-piperazine, which has thestructural formula

[1021] was prepared as follows. A mixture of3R-methyl-1-(4-nitro-phenyl)-piperazine (124 mg, 0.56 mmol), sodiumformate (93 mg, 1.37 mmol), formic acid (1.5 mL), and formalin (1.5 mL)was stirred at 80° C. overnight, cooled, poured into ice/water, andextracted with CHCl₃. The organic layer washed with brine, dried overNa₂SO₄, and concentrated to give 116 mg (71% yield) of yellow crystals,which were used without further purification.

[1022]¹H NMR (CDCl₃): δ 8.12 (2H, d, J=9.4 Hz), 6.82 (2H, d, J=9.4 Hz),3.76 (1H, d, J=12.4 Hz), 3.67 (1H, d, J=12.4 Hz), 3.14 (1H, ddd, J=12.4,11.7, 1.5 Hz), 2.90 (1H, d, J=11.7 Hz), 2.74 (1H, dd, J=11.7, 10.9 Hz),2.40 (1H, m), 2.34 (3H, s), 2.22 (1H, m), 1.16 (3H, d, J=6.3 Hz).

[1023] 4-(3R,4-Dimethyl-piperazin-1-yl)-aniline, which has thestructural formula

[1024] was made as follows. A mixture of1,2R-dimethyl-4-(4-nitro-phenyl)-piperazine (168 mg, 0.71 mmol), 10%Pd/C (30 mg), and MeOH (10 mL) was stirred under hydrogen for 1.5 hours.The catalyst was filtered off and the filtrate concentrated in vacuo toprovide a cloudy yellow oil, which was used without furtherpurification.

[1025]¹H NMR (CDCl₃): δ 6.91 (2H, d, J=8.8 Hz), 6.75 (2H, d, J=8.8 Hz),3.66-3.32 (4H, m), 3.05-2.89 (2H, m), 2.63-2.48 (2H, m), 2.44-2.36 (1H,m), 2.44 (3H, s), 1.22 (3H, d, J=6.1 Hz).

[1026] 4-(4-Isothiocyanato-phenyl)-1,2R-dimethyl-piperazine, which hasthe structural formula

[1027] was prepared as follows. To4-(3R,4-dimethyl-piperazin-1-yl)-aniline (0.71 mmol) in THF (15 mL) at−35° C. was added in succession Et₃N (0.20 mL, 1.43 mmol) andthiophosgene (58 μl, 0.75 mmol) dropwise. The solvent was evaporated andthe residue partitioned with CHCl₃ and water. The organic layer wasdried with Na₂SO₄ and concentrated to furnish a brown powder, 184 mg,which contained trace Et₃N by NMR, but was sufficient for use withoutfurther purification.

[1028]¹H NMR (CDCl₃): δ 7.12 (2H, d, J=9.1 Hz), 6.82 (2H, d, J=9.1 Hz),3.58-3.46 (2H, m), 3.13-3.03 (2H, m), 2.89-2.75 (1H, m), 2.65-2.41 (2H,m), 2.49 (3H, s), 1.27 (3H, d, J=6.3 Hz).

[1029] The title compound was prepared in a manner analogous to thatused in Example C(1).4-(4-Isothiocyanato-phenyl)-1,2R-dimethyl-piperazine and2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided ayellow powder in 57% yield, mp 115-118° C.

[1030]¹H NMR (DMSO-d₆): δ 10.65 (1H, bs), 8.15 (2H, bs), 7.62-7.50 (1H,m), 7.35 (2H, d, J=9.0 Hz), 7.23 (1H, d, J=7.7 Hz), 7.20 (1H, d, J=8.0Hz), 6.97 (2H, d, J=9.0 Hz), 3.59-3.49 (2H, m), 3.34 (3H, s), 2.90-2.72(2H, m), 2.40 (1H, t, J=10.9 Hz), 2.28-2.05 (2H, m), 1.09 (3H, d, J=6.2Hz).

[1031] HRFABMS (MH⁺): Calcd.: 444.1670. Found: 444.1656.

[1032] Anal. Calcd. for C₂₂H₂₃N₅OSF₂. 0.8H₂O.0.6t-BuOH: C, 58.33; H,6.14; N, 13.94; S, 6.38. Found: C, 58.38; H, 5.92; N, 13.89; S, 6.33.

Example C(125) {4-Amino-2-[4-(3S,4-dimethyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(2,6-difluoro-phenyl)-methanone

[1033]

[1034] 4-(4-Isothiocyanato-phenyl)-1,2S-dimethyl-piperazine, which hasthe structural formula

[1035] was prepared according to the route employed for its enantiomer,4-(4-isothiocyanato-phenyl)-1,2R-dimethyl-piperazine for Example C(24).The resultant yellow powder displayed a comparable NMR spectrum and wasused without further purification.

[1036] The title compound was prepared in a manner analogous to thatused in Example C(1).4-(4-Isothiocyanato-phenyl)-1,2S-dimethyl-piperazine and2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided ayellow powder in 77% yield, mp 110-116° C.

[1037]¹H NMR (DMSO-d₆): δ 10.65 (1H, bs), 8.15 (2H, bs), 7.62-7.50 (1H,m), 7.35 (2H, d, J=9.0 Hz), 7.23 (1H, d, J=7.7 Hz), 7.20 (1H, d, J=8.0Hz), 6.97 (2H, d, J=9.0 Hz), 3.59-3.49 (2H, m), 3.34 (3H, s), 2.90-2.72(2H, m), 2.40 (1H, t, J=10.9 Hz), 2.28-2.05 (2H, m), 1.09 (3H, d, J=6.2Hz).

[1038] IR (KBr): 3386, 3274, 3168, 2970, 2807, 1620, 1589, 1547, 1517,1464, 1429, 1238, 1001 cm⁻¹.

[1039] HRFABMS (MH⁺): Calcd.: 444.1670. Found: 444.1659.

[1040] Anal. Calcd. for C₂₂H₂₃N₅OSF₂.0.7H₂O.0.2t-BuOH: C, 58.15; H,5.65; N, 14.87; S, 6.81. Found: C, 58.06; H, 5.61; N, 14.58; S, 6.90.

Example C(126)(4-Amino-2-{4-[(3-dimethylamino-propyl)-methyl-amino]-phenylamino}-thiazol-5-yl)-(2,6-difluoro-phenyl)-methanone

[1041]

[1042] N-(4-Nitrophenyl)-N,N′,N′-trimethyl-propane-1,3-diamine, whichhas the structural formula

[1043] was first prepared in a manner analogous to tert-butyl[methyl-(4-nitro-phenyl)-amino]-acetate for Example C(103).4-Fluoronitrobenzene and N,N,N′-trimethyl-propanediamine gave a yellowoil, which was heated up to 280° C. at 1 torr to remove startingmaterials, furnishing an orange oil, 4.26 g (85% crude yield), which wasused without any further purification.

[1044]¹H NMR (CDCl₃): δ 8.10 (2H, ddd, J=9.5, 8.2, 5.3 Hz), 6.64 (2H,ddd, J=9.5, 8.2, 5.3 Hz), 3.50 (2H, t, J=7.2 Hz), 3.08 (3H, s), 2.07(3H, t, J=6.8 Hz), 2.23 (6H, s), 1.72-1.82 (2H, m).

[1045] N-(4-Aminophenyl)-N,N′,N′-propane-1,3-diamine, which has thestructural formula

[1046] was prepared as follows. A mixture ofN-(4-nitrophenyl)-N,N′,N′-trimethyl-propane-1,3-diamine (1.72 g, 7.25mmol), tin(II) chloride dihydrate (8.05 g, 36.2 mmol), dioxane (25 mL),and ethanol (5 mL) was heated at reflux for 3.5 hours, then allowed tocool. To the resultant mixture was added sat aq. Na₂CO₃ until no gasevolution was observed. Celite was added to ease subsequent filtering.The solids were rinsed with MeOH, and the filtrate was concentratedunder reduced pressure and extracted with 10% MeOH/CHCl₃ (4×). Thecombined extracts were washed with brine, dried over Na₂SO₄, andevaporated to give a black oil, which was purified via columnchromatography with alumina (neutral, activity 1) and 1% MeOH/CH₂Cl₂ aseluant to afford 0.39 g (26%) of a darkening brown oil that was usedwithout further purification.

[1047]¹H NMR (CDCl₃): δ 6.67 (4H, dd, J=9.0, 8.6 Hz), 3.22 (2H, t, J=7.2Hz), 2.82 (3H, s), 2.31 (2H, t, J=7.5 Hz), 2.23 (6H, s), 1.70 (2H, p,J=7.4 Hz).

[1048]N-(4-Isothiocyanato-phenyl)-N,N′,N′-trimethyl-propane-1,3-diamine, whichhas the structural formula

[1049] was prepared in a manner analogous to4-(4-isothiocyanato-phenyl)-1,2R-dimethyl-piperazine for Example C(124).N-(4-Aminophenyl)-N,N′,N′-propane-1,3-diamine provided a black oil in86% crude yield, which was used without further purification.

[1050]¹H NMR (CDCl₃): δ 7.09 (211, d, J=9.0 Hz), 6.59 (2H, d, J=9.0 Hz),3.38 (2H, J=7.2 Hz), 2.94 (3H, s), 2.36 (2H, t, J=7.2 Hz), 2.29 (6H, s),1.78 (2H, p, J=7.2 Hz). IR (KBr): 2127, 1605, 1514, 1379 cm⁻¹.

[1051] The title compound was prepared in a manner like that describedfor Example C(1).N-(4-Isothiocyanato-phenyl)-N,N′,N′-trimethyl-propane-1,3-diamine and2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) afforded abrown oil, which was purified via flash column chromatography with astepwise gradient of 7-14% MeOH/CHCl₃ and precipitated from CH₂Cl₂/hexto provide an amorphous yellow solid in 51% yield, mp 115-120° C.(decomp).

[1052]¹H NMR (DMSO-d₆): δ 10.50 (1H, bs), 8.05 (2H, bs), 7.50 (1H, ddd,J=15.3, 8.4, 6.7 Hz), 7.10-7.35 (4H, m), 6.68 (2H, d, J=9.1 Hz), 2.84(3H, s), 2.27 (2H, t, J=7.2 Hz), 2.16 (6H, s), 1.61 (2H, p, J=7.3 Hz).

[1053] IR (KBr): 3393, 3279, 3165, 2951, 1619, 1545, 1524, 1462, 1436cm⁻¹.

[1054] HRFABMS: Calcd. for C₂₂H₂₆F₂N₅OS (MH⁺): 446.1826. Found:446.1810.

[1055] Anal. Calcd. for C₂₁H₂₃F₂N₅OS.0.8H₂O.0.4C₆H₁₄: C, 59.28; H, 6.56;N, 14.16; S, 6.49. Found: C, 59.37; H, 6.31; N, 13.76; S, 6.26.

Example C(127)(2,6-Difluoro-phenyl)-{2-[4-(4-pyridin-4-yl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-methanone

[1056]

[1057] 1-(4-Nitro-phenyl)-4-pyridin-4-yl-piperazine, which has thestructural formula

[1058] was prepared in a manner analogous to tert-butyl[methyl-(4-nitro-phenyl)-amino]-acetate for Example C(103).4-Fluoronitrobenzene and 1-(4-pyridyl)piperazine (Ratous et. al., J.Med. Chem., vol. 8 (1965), pp. 104-107) gave a brown powder in 27%yield, which was used without further purification.

[1059]¹H NMR (CD₃OD): δ 8.20 (2H, d, J=5.0 Hz), 8.08 (2H, d, J=9.4 Hz),7.04 (2H, d, J=9.5 Hz), 3.62-3.68 (4H, m), 3.50-3.56 (4H, m).

[1060] 4-(4-Pyridin-4-yl-piperazin-1-yl)-aniline, which has thestructural formula

[1061] was prepared in a manner analogous to4-(4-methyl-piperazin-1-yl)-aniline for Example C(70).1-(4-Nitro-phenyl)-4-pyridin-4-yl-piperazine afforded a brown powder in100% crude yield, which was used without further purification.

[1062]¹H NMR (CD₃OD): δ 8.16 (2H, d, J=6.7 Hz), 6.90 (4H, bd, J=8.9 Hz),6.74 (2H, d, J=6.6 Hz), 3.56 (4H, dd, J=5.3, 5.0 Hz), 3.14 (4H, dd,J=5.0, 4.2 Hz).

[1063] 1-(4-Isothiocyanato-phenyl)-4-pyridin-4-yl-piperazine, which hasthe structural formula

[1064] was prepared as follows. To a solution of4-(4-pyridin-4-yl-piperazin-1-yl)-aniline (2.00 g, 7.86 mmol) in 10% aqHCl (10 mL) was added thiophosgene (720 μL, 9.43 mmol). After 0.5 hour,the resultant yellow precipitate was filtered off, washed with sat aqNaHCO₃ and H₂O, and dried under high vacuum to give 1.9 g (82% yield) ofa yellow powder, which was used without further purification.

[1065]¹H NMR (DMSO-d₆): δ 6.73 (4H, d, J=8.8 Hz), 6.51 (4H, d, J=8.8Hz), 3.32 (4H, bs), 3.29 (4H, bs).

[1066] The title compound was prepared in a manner like that describedfor Example C(1). 1-(4-Isothiocyanato-phenyl)-4-pyridin-4-yl-piperazineand 2-bromo-2′,6′-difluoroacetophenone (from Example C(79)) provided,after recrystallization with trace DMSO in MeOH/CHCl₃, a pale tan powderin 30% yield, mp 155-157° C.

[1067]¹H NMR (DMSO-d₆): δ 8.16 (2H, d, J=6.0 Hz), 8.04 (1H, bs),7.40-7.52 (1H, m), 7.32 (2H, d, J=8.7 Hz), 7.15 (2H, t, J=7.7 Hz), 6.96(2H, d, J=9.0 Hz), 6.85 (2H, d, J=5.5 Hz), 3.60 (4H, bs).

[1068] HRFABMS: Calcd. for C₂₅H₂₃F₂N₆OS (MH⁺): 493.1622. Found:493.1606.

[1069] Anal. Calcd. for C₂₅H₂₂F₂N₆OS.0.7MeOH.0.1CHCl₃.0.1DMSO: C, 58.40;H, 4.81; N, 15.72; S, 6.60. Found: C, 58.38; H, 4.50; N, 15.37; S, 7.00.

Example C(128){4-Amino-2-[4-(1-methyl-[1,4]-diazepan-4-yl)-phenylamino]-thiazol-5-yl}-(2,6-difluoro-phenyl)-methanone

[1070]

[1071] 1-Methyl-4-(4-nitro-phenyl)-[1,4]diazepane, which has thestructural formula

[1072] was prepared in a manner analogous to tert-butyl[methyl-(4-nitro-phenyl)-amino]-acetate for Example C(103).1-Methyl-homopiperazine provided a yellow powder in 93% yield, which wasused without further purification.

[1073]¹H NMR (CDCl₃): δ 8.12 (2H, d, J=9.5 Hz), 6.64 (2H, d, J=9.5 Hz),3.56-3.70 (4H, m), 2.74 (2H, dd, J=4.9, 3.3 Hz), 2.58 (2H, dd, J=5.6,5.4 Hz), 2.40 (3H, s), 2.00-2.08 (2H, m).

[1074] 4-(4-Methyl-[1,4]diazepan-1-yl)-aniline, which has the structuralformula

[1075] was prepared in a manner analogous to4-(4-methyl-piperazin-1-yl)-aniline for Example C(70).1-Methyl-4-(4-nitro-phenyl)-[1,4]diazepane furnished a purple oil in100% crude yield, which was used immediately without furtherpurification.

[1076]¹H NMR (CDCl₃): δ 6.68 (2H, d, J=12.2 Hz), 6.60 (2H, d, J=6.8 Hz),3.52 (2H, dd, J=4.8, 4.7 Hz), 3.43 (2H, t, J=6.3 Hz), 2.71 (2H, dd,J=4.9, 4.7 Hz), 2.58 (2H, dd, J=5.5, 5.4 Hz), 2.38 (3H, s), 1.95-2.04(1H, m).

[1077] 1-(4-Isothiocyanato-phenyl)-4-methyl-[1,4]diazepane, which hasthe structural formula

[1078] was prepared in a manner analogous to1-(4-isothiocyanato-phenyl)-1H-imidazole for Example C(41).4-(4-Methyl-[1,4]diazepan-1-yl)-aniline gave a crude product that wasextracted with CHCl₃ to eventually afford a black oil in 85% crudeyield. This material was used immediately without any furtherpurification.

[1079]¹H NMR (CDCl₃): δ 7.02 (2H, d, J=9.0 Hz), 6.56 (2H, d, J=9.0 Hz),3.54 (2H, dd, J=4.8, 4.8 Hz), 3.45 (2H, t, J=6.3 Hz), 2.67 (2H, dd,J=4.9, 4.8 Hz), 2.53 (2H, dd, J=5.6, 5.4 Hz), 2.36 (3H, s), 1.97 (2H, p,J=5.7 Hz).

[1080] The title compound was prepared in a manner like that describedfor Example C(1). 1-(4-Isothiocyanato-phenyl)-4-methyl-[1,4]diazepaneand 2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided,after crystallization from boiling EtOH, a light-tan powder in 26%yield, mp 138-140° C.

[1081]¹H NMR (DMSO-d₆): δ 8.05 (1H, s), 7.42-7.52 (1H, m), 7.10-7.22(4H, m), 6.64 (2H, d, J=9.1 Hz), 3.36-3.52 (4H, m), 2.58 (2H, dd, J=4.8,4.7 Hz), 2.42 (2H, dd, J=5.6, 5.4 Hz), 2.25 (3H, s), 1.82-1.92 (2H, m).

[1082] HRFABMS: Calcd. for C₂₂H₂₄F₂N₅OS (MH⁺): 444.1670. Found:444.1656.

[1083] Anal. Calcd. for C₂₂H₂₃F₂N₅OS.0.5H₂O.0.8EtOH: C, 57.92; H, 5.93;N, 14.31; S, 6.55. Found: C, 58.05; H, 5.69; N, 14.15; S, 6.55.

Example C(129)3-({4-[4-Amino-5-(2,6-difluorobenzoyl)-thiazol-2-yl-amino]-phenyl}-methylamino)-propionitrile

[1084]

[1085] 3-[Methyl-(4-nitro-phenyl)-amino]-propionitrile, which has thestructural formula

[1086] was prepared as follows. Benzyltrimethylammonium hydroxide (7.23mL of a 40% solution in MeOH) was added to a suspension ofN-methyl-4-nitroaniline (5.00 g, 32.9 mmol) and acrylonitrile (7.23 mL)in dioxane (80 mL). The resultant solution was heated at 55° C. for 3.5hours, then poured into water, and extracted with 20% isopropanol inchloroform. The separated organic layer was washed with water, driedover K₂CO₃, and concentrated to a suspension of yellow solid, which wasdiluted with ether. The solid was filtered off and dried under vacuum toobtain 6.15 g (91% yield) of yellow solid, which was used withoutfurther purification.

[1087]¹H NMR (CDCl₃): δ 8.17 (2H, d, J=9.4 Hz), 6.66 (2H, d, J=9.4 Hz),3.82 (2H, t, J=6.7 Hz), 3.19 (3H, s), 2.66 (2H, t, J=6.7 Hz).

[1088] 3-[(4-Amino-phenyl)-methyl-amino]-propionitrile, which has thestructural formula

[1089] was prepared in a manner analogous to4-(3S,4-dimethyl-piperazin-1-yl)-phenylamine for Example C(134).3-[Methyl-(4-nitro-phenyl)-amino]-propionitrile gave a brown oil in 100%yield, which was used without further purification.

[1090]¹H NMR (CDCl₃): δ 6.68 (4H, s), 3.57 (2H, t, J=7.0 Hz), 2.90 (3H,s), 2.51 (2H, t, J=7.0 Hz).

[1091] 3-[(4-Isothiocyanato-phenyl)-methyl-amino]-propionitrile, whichhas the structural formula

[1092] was prepared in a manner analogous to4-(4-isothiocyanato-phenyl)-1,2 S-dimethyl-piperazine for ExampleC(134). 3-[(4-Amino-phenyl)-methyl-amino]-propionitrile gave a brownsolid in 95% yield, which was used without further purification.

[1093]¹H NMR (CDCl₃): δ 7.15 (2H, d, J=9.1 Hz), 6.62 (2H, d, J=9.1 Hz),3.72 (2H, t, J=6.8 Hz), 3.05 (3H, s), 2.58 (2H, t, J=6.8 Hz).

[1094] The title compound was prepared in a manner analogous to thatused in Example C(1).3-[4-(4-Isothiocyanato-phenyl)-methylamino]-propionitrile and2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided anamorphous yellow powder in 66% yield, mp 120-130° C. (decomp).

[1095]¹H NMR (DMSO-d₆): δ 10.52 (1H, bs), 8.10 (2H, bs), 7.49 (1H, ddd,J=15.3, 8.2, 6.7 Hz), 7.26 (2H, bd, J=8.2 Hz), 7.15 (2H, dd, J=8.1, 7.7Hz), 6.76 (2H, d, J=9.1 Hz), 3.63 (2H, t, J=6.7 Hz), 2.91 (3H, s), 2.69(2H, t, J=6.7 Hz).

[1096] IR (KBr): 3417, 3309, 1618, 1548, 1523, 1463, 1436, 1376, 1356,1234, 1001 cm⁻¹.

[1097] HRFABMS Calcd. for C₂₀H₁₇N₅OSF₂Na (M+Na⁺): 436.1020. Found:436.1030.

[1098] Anal. Calcd. for C₂₀H₁₇N₅OSF₂.0.2H₂O.0.45t-BuOH: C, 58.13; H,4.90; N, 15.55; S, 7.12. Found: C, 57.88; H, 4.79; N, 15.16; S, 6.95.

Example C(130)2-[4-Amino-2-(4-nitro-phenylamino)-thiazole-5-carbonyl]-phenyl Benzoate

[1099]

[1100] The title compound was prepared essentially as described forExample C(1). In addition, two other reaction products were isolatedafter flash column chromatography and identified: characteristics for(Z)- and(E)-4-(2-hydroxy-phenyl)-3-(4-nitro-phenyl)-3H-thiazol-2-ylidene-cyanamidefollow below. 4-Nitro-phenyl isothiocyanate and2′-benzoyloxy-2-bromo-acetophenone provided title compound as a yellowsolid, mp 258-260° C.

[1101]¹H NMR (DMSO-d₆): δ 11.35 (1H, s), 8.23 (2H, d, J=9.3 Hz),7.98-8.04 (4H, m), 7.85 (2H, d, J=9.2 Hz), 7.35-7.67 (1H, m), 7.52-7.63(4H, m), 7.39-7.45 (2H, m).

[1102]¹³C NMR(MeOH-d₄): δ 181.5, 166.4, 164.4, 147.2, 145.8, 142.0,135.2, 134.3, 131.2, 130.0, 129.3, 129.2, 128.3, 126.5, 125.6, 123.9,118.3.

[1103] Anal. Calcd. for C₂₃H₁₆N₄O₅S: C, 59.99; H, 3.50; N, 12.17; S,6.96. Found: C, 58.25; H, 3.54; N, 11.77; S, 6.94.

[1104] Earlier-eluting component,(Z)-4-(2-hydroxy-phenyl)-3-(4-nitro-phenyl)-3H-thiazol-2-ylidene-cyanamide,which has the structural formula

[1105] was isolated as a yellow amorphous solid.

[1106]¹H NMR (DMSO-d₆): δ 9.79 (1H, s), 8.18 (2H, d, J=9.0 Hz), 7.55(2H, d, J=9.0 Hz), 7.26 (1H, dd, J=7.5, 1.5 Hz), 7.17 (1H, ddd, J=7.5,7.4, 1.5 Hz), 7.05 (1H, s), 6.79 (1H, dd, J=7.6, 7.4 Hz), 6.65 (11H, d,8.2 Hz).

[1107]¹³C NMR (MeOH-d₄): δ 176.8, 157.9, 150.3, 143.6, 141.7, 134.4,134.2, 132.0, 126.0, 122.1, 119.5, 119.3, 117.9, 107.3.

[1108] HRFABMS: Calcd. for C₁₆H₁₀N₄O₃S (MH⁺): 339.0552. Found: 339.0550.

[1109] A later-eluting component,(E)-4-(2-hydroxy-phenyl)-3-(4-nitro-phenyl)-3H-thiazol-2-ylidene-cyanamide,which has the structural formula

[1110] was isolated as a yellow amorphous solid.

[1111]¹H NMR (DMSO-d₆): δ 13.2(11H, s), 8.25 (2H, d, J=9.2 Hz), 7.75(1H, dd, J=7.8, 1.5 Hz), 7.55 (1H, ddd, J=8.6, 7.5, 1.1 Hz), 7.41 (1H,ddd, J=8.6, 7.5, 1.1 Hz), 7.25 (1H, dd, J=8.1, 1.0 Hz), 7.13 (1H, d, 9.2Hz), 7.01 (1H, s).

[1112]¹³C NMR (MeOH-d₄): δ 174.8, 162.1, 152.2, 143.6, 134.0, 131.4,129.9, 126.4, 126.2, 122.0, 121.5, 117.8, 105.6.

[1113] ESIMS: Calcd. for C₁₆H₁₀N₄O₃S (MH⁺): 339. Found: 339.

Example C(131)(4-Amino-2-{4-[4-(2,2,2-trifluoro-ethyl)-piperazin-1-yl]-phenylamino}-thiazol-5-yl)-(2,6-difluoro-phenyl)-methanone

[1114]

[1115] 1-(4-Nitro-phenyl)-4-(2,2,2-trifluoro-ethyl)-piperazine, whichhas the structural formula

[1116] was first prepared in a manner analogous to tert-butyl[methyl-(4-nitro-phenyl)-amino]-acetate for Example C(103).1-(4-Nitro-phenyl)-piperazine and 1,1,1-trifluoro-2-iodo-ethane gave ayellow-orange solid in 33% crude yield.

[1117]¹H NMR (CDCl3): δ 8.13 (2H, d, J=9.2 Hz), 6.82 (2H, d, J=9.2 Hz),3.51-3.38 (4H, m), 3.10-2.99 (2H, m), 2.87-2.77 (4H, m).

[1118] 4-[4-(2,2,2-Trifluoro-ethyl)-piperazin-1-yl]-aniline, which hasthe structural formula

[1119] was next prepared in a manner analogous to4-(4-methyl-piperazin-1-yl)-aniline for Example C(70).1-(4-Nitro-phenyl)-4-(2,2,2-trifluoro-ethyl)-piperazine gave apale-brown solid in 100% crude yield.

[1120]¹H NMR (CDCl3): δ 6.83 (2H, d, J=8.8 Hz), 6.68 (2H, d, J=8.8 Hz),3.40 (2H, bs), 3.11-3.06 (6H, m), 2.86 (4H, dd, J 5.1, 4.7 Hz).

[1121] 1-(4-Isothiocyanato-phenyl)-4-(2,2,2-trifluoro-ethyl)-piperazine,which has the structural formula

[1122] was prepared in a manner analogous to1-(4-isothiocyanato-phenyl)-4-methyl-piperazine for Example C(70) from4-[4-(2,2,2-trifluoro-ethyl)-piperazin-1-yl]-aniline, providing a brownpowder in 89% yield.

[1123]¹H NMR (CDCl3): δ 7.15 (2H, d, J=9.1 Hz), 6.85 (2H, d, J=9.0 Hz),3.25 (4H, dd, J=4.9, 5.2 Hz), 3.05 (2H, q, J=9.5 Hz), 2.86 (4H, dd, J5.1, 4.8 Hz).

[1124] The title compound was prepared in a manner like that describedfor Example C(1).1-(4-isothiocyanato-phenyl)-4-(2,2,2-trifluoro-ethyl)-piperazine and2-bromo-2′,6′-difluoroacetophenone (from Example C(79)) provided, afterpurification via column chromatography with 5% MeOH/CHCl3 as eluant, ayellow powder in 63% yield, mp 99-102° C.

[1125]¹H NMR (DMSO-d₆): δ 8.12 (1H, bs), 7.58-7.46 (1H, m), 7.30 (2H,bd, J=7.4 Hz), 7.18 (2H, dd, J=7.8, 7,7 Hz), 6.92 (2H, d, J=8.9 Hz),3.24 (2H, q, J=10.3 Hz), 3.12 (4H, dd, J=4.1, 5.0 Hz), 2.76 (4H, bd,J=4.6 Hz).

[1126] IR (KBr): 3394, 3276, 3178, 3058, 2954, 2829, 1617, 1588, 1547,1462, 1426, 1231 cm⁻¹.

[1127] Anal. Calcd. for C₂₂H₂OF₂N₅OS.0.15CHCl₃: C, 51.62; H, 3.94; N,13.59; S, 6.22. Found: C, 51.68; H, 3.93; N, 13.39; S, 6.03.

Example C(131)(4-Amino-2-{4-[4-(2,2,2-trifluoroethyl)-piperazin-1-yl]-phenylamino}-thiazol-5-yl)-(2,6-difluoro-phenyl)-methanone

[1128]

[1129] 1-(4-Nitro-phenyl)-4-(2,2,2-trifluoro-ethyl)-piperazine, whichhas the structural formula

[1130] was first prepared in a manner analogous to tert-butyl[methyl-(4-nitro-phenyl)-amino]-acetate for Example C(103).1-(4-Nitro-phenyl)-piperazine and 1,1,1-trifluoro-2-iodo-ethane gave ayellow-orange solid in 33% crude yield.

[1131]¹H NMR (CDCl₃): δ 8.13 (2H, d, J=9.2 Hz), 6.82 (2H, d, J=9.2 Hz),3.51-3.38 (4H, m), 3.10-2.99 (2H, m), 2.87-2.77 (4H, m).

[1132] 4-[4-(2,2,2-Trifluoro-ethyl)-piperazin-1-yl]-aniline, which hasthe structural formula

[1133] was next prepared in a manner analogous to4-(4-methyl-piperazin-1-yl)-aniline for Example C(70).1-(4-Nitro-phenyl)-4-(2,2,2-trifluoro-ethyl)-piperazine gave apale-brown solid in 100% crude yield.

[1134]¹H NMR (CDCl₃): δ 6.83 (2H, d, J=8.8 Hz), 6.68 (2H, d, J=8.8 Hz),3.40 (2H, bs), 3.11-3.06 (6H, m), 2.86 (4H, dd, J=5.1, 4.7 Hz).

[1135] 1-(4-Isothiocyanato-phenyl)-4-(2,2,2-trifluoro-ethyl)-piperazine,which has the structural formula

[1136] was prepared in a manner analogous to1-(4-isothiocyanato-phenyl)-4-methyl-piperazine for Example C(70) from4-[4-(2,2,2-trifluoro-ethyl)-piperazin-1-yl]-aniline, providing a brownpowder in 89% yield.

[1137]¹H NMR (CDCl₃): δ 7.15 (2H, d, J=9.1 Hz), 6.85 (2H, d, J=9.0 Hz),3.25 (4H, dd, J=4.9, 5.2 Hz), 3.05 (2H, q, J=9.5 Hz), 2.86 (4H, dd,J=5.1, 4.8 Hz).

[1138] The title compound was prepared in a manner like that describedfor Example C(1).1-(4-isothiocyanato-phenyl)-4-(2,2,2-trifluoro-ethyl)-piperazine and2-bromo-2′,6′-difluoroacetophenone (from Example C(79)) provided, afterpurification via column chromatography with 5% MeOH/CHCl3 as eluant, ayellow powder in 63% yield, mp 99-102° C.

[1139]¹H NMR (DMSO-d₆): δ 8.12 (1H, bs), 7.58-7.46 (1H, m), 7.30 (2H,bd, J=7.4 Hz), 7.18 (2H, dd, J=7.8, 7,7 Hz), 6.92 (2H, d, J=8.9 Hz),3.24 (2H, q, J=10.3 Hz), 3.12 (4H, dd, J=4.1, 5.0 Hz), 2.76 (4H, bd,J=4.6 Hz).

[1140] IR (KBr): 3394, 3276, 3178, 3058, 2954, 2829, 1617, 1588, 1547,1462, 1426, 1231 cm⁻¹.

[1141] Anal. Calcd. for C₂₂H₂OF₂N₅OS.0.15CHCl₃: C, 51.62; H, 3.94; N,13.59; S, 6.22. Found: C, 51.68; H, 3.93; N, 13.39; S, 6.03.

Example D(1)(3-Amino-phenyl)-(4-amino-2-phenylamino-thiazol-5-yl)-methanone

[1142]

[1143] A mixture of the title compound from Example A(1)((4-amino-2-phenylamino-thiazol-5-yl)-(3-nitrophenyl)-methanone, 520 mg,1.53 mmol) and 10% palladium on carbon (80 mg) in THF (10 mL) wasstirred under a hydrogen atmosphere overnight. The catalyst was filteredoff and the filtrate concentrated in vacuo to give 470 mg of a crudesolid that recrystallized from ethyl acetate/benzene to provide 100 mg(19% yield) of light yellow powder, mp 162-164° C.

[1144]¹H NMR (DMSO-d₆): δ 10.75 (1H, s), 8.42 (2H, bs), 8.15 (2H, bs),7.60 (2H, d, J=7.8 Hz), 7.34 (2H, d, J=7.8 Hz), 7.23 (1H, t, J=7.8 Hz),7.14 (1H, s), 7.07 (11H, d, J=7.8 Hz), 7.05 (1H, t, J=7.8 Hz), 6.91 (1H,d, J=7.8 Hz).

[1145] FABMS (MH⁺): 311.

[1146] Anal. Calcd. for C₁₆H₁₄N₄OS.H₂O.C₆H₆: C, 59.30; H, 4.98; N,16.66; S, 9.54. Found: C, 59.02; H, 4.61; N, 16.34; S, 9.25.

Example D(2)(4-Amino-phenyl)-(4-amino-2-phenylamino-thiazol-5-yl)-methanone

[1147]

[1148] The title compound was prepared in a manner like that describedfor Example D(1). Catalytic reduction of the title compound of ExampleA(2) ((4-nitro-phenyl)-(4-amino-2-phenylamino-thiazol-5-yl)-methanone)provided, after recrystallization from ethanol, 410 mg (90% yield) ofred amorphous powder, mp>300° C.

[1149]¹H NMR (DMSO-d₆): δ 10.85 (1H, bs), 8.44-8.20 (2H, bs), 8.36 (1H,d, J=8.7 Hz), 8.17 (11H, d, J=8.7 Hz), 7.89 (1H, d, J=15.9 Hz), 7.86(1H, d, J=15.9 Hz), 7.62 (2H, d, J=7.8 Hz), 7.37 (2H, t, J=7.8 Hz), 7.09(1H, t, J=7.8 Hz).

[1150] FABMS (MH⁺): 311.

[1151] Anal. Calcd. for C₁₆H₁₄N₄OS.0.5H₂O: C, 60.17; H, 4.73; N, 17.54;S, 10.04. Found: C, 60.09; H, 4.73; N, 17.58; S, 9.93.

Example D(3)[4-Amino-2-(4-dimethylamino-phenylamino)-thiazol-5-yl]-(2-amino-phenyl)-methanone

[1152]

[1153] The title compound was prepared essentially as described forExample D(1). Catalytic reduction of the title compound of Example C(4)gave 26 mg (30% yield) of an amorphous solid.

[1154]¹H NMR (DMSO-d₆): δ 10.38 (1H, s), 8.06 (2H, bs), 7.31 (2H, d,J=9.0 Hz), 7.30 (1H, d, J=7.5 Hz), 7.08 (1H, t, J=7.5 Hz), 6.72 (2H, d,J=9.0 Hz), 6.68 (1H, d, J=7.5 Hz), 6.51 (1H, t, J=7.5 Hz), 5.75 (2H, s),2.88 (6H, s).

[1155] FABMS (MH⁺): 354.

[1156] Anal. Calcd. for C₁₈H₁₉N₅OS.0.5H₂O.0.3MeOH: C, 59.07; H, 5.74; N,18.82; S, 8.62. Found: C, 59.24; H, 5.56; N, 18.51; S, 8.36.

Example D(4)[4-Amino-2-(4-amino-phenylamino)-thiazol-5-yl]-phenyl-methanone

[1157]

[1158] The title compound was prepared in a manner similar to thatdescribed for Example D(1). Catalytic reduction of the title compoundfrom Example A(8) (i.e.,[4-amino-2-(4-nitro-phenylamino)-thiazol-5-yl]-phenyl-methanone, 450 mg,1.32 mmol) gave, after recrystallization from ethanol, 120 mg (29%yield) of orange powder, mp 167-169° C.

[1159]¹H NMR (DMSO-d₆): δ 10.38 (1H, s), 8.15 (2H, bs), 7.64-7.55 (2H,m), 7.47-7.38 (3H, m), 7.10 (2H, d, J=8.6 Hz), 6.55 (2H, d, J=8.6 Hz),5.20 (2H, bs).

[1160] FABMS (MH⁺): 311.

[1161] Anal. Calcd. for C₁₆H₁₄N₄OS.H₂O: C, 56.96; H, 5.08; N, 16.61; S,9.50. Found: C, 56.94; H, 5.07; N, 16.60; S, 9.64.

Example D(5)4-[4-Amino-5-(3-amino-5-amino-thiophene-2-carbonyl)-thiazol-2-ylamino]-benzenesulfonamide

[1162]

[1163] The title compound was prepared in a manner analogous to thatused in Example D(1). The title compound of Example C(95) washydrogenated and recrystallized from EtOH to provide a brown powder in96% yield, mp 268-271° C.

[1164]¹H NMR (DMSO-d₆): δ 10.97 (1H, s), 7.91 (2H, s), 7.82 (2H, d,J=9.1 Hz), 7.78 (2H, d, J=9.1 Hz), 7.28 (2H, s), 6.43 (2H, s), 5.81 (1H,s), 2.34 (3H, s).

[1165] FABMS (MH⁺): 410.

[1166] Anal. Calcd. for C₁₅H₁₅N₅O₃S₃.0.1H₂O.0.3EtOH: C, 44.07; H, 4.03;N, 16.47; S, 22.63. Found: C, 44.23; H, 3.93; N, 16.07; S, 23.01.

Example E(1) 4-[4-Amino-5-(2-nitro-benzoyl)-thiazol-2-ylamino]-benzoicAcid

[1167]

[1168] To a suspension of the title compound of Example A(5) (i.e.,ethyl 4-[4-amino-5-(2-nitro-benzoyl)-thiazol-2-ylamino]-benzoate, 950mg, 2.3 mmol), in methanol (15 mL) was added 3N NaOH (10 mL). After 30minutes, the mixture was acidified to a pH of 4 with 1N HCl, whereupon ayellow precipitate formed. The mixture was diluted with water (100 mL).The solid was filtered off and rinsed with water. Recrystallization fromethanol provided 672 mg (76% yield) of yellow crystals, mp 289-292° C.

[1169]¹H NMR (DMSO-d₆): δ 12.75 (1H, s), 11.13 (1H, s), 8.12 (2H, bs),8.08 (1H, d, J=7.8 Hz), 7.91 (2H, d, J=8.7 Hz), 7.82 (1H, td, J=8.4, 0.9Hz), 7.78-7.68 (4H, m).

[1170] FABMS (MH⁺): 385.

[1171] Anal. Calcd. for C₁₉H₁₈N₄O₃S: C, 53.12; H, 3.15; N, 14.58; S,8.34. Found: C, 53.29; H, 3.25; N, 14.31; S, 8.11.

Example E(2)4-[4-Amino-2-(4-sulfamoyl-phenylamino)-thiazole-5-carbonyl]-benzoic Acid

[1172]

[1173] To a suspension of ethyl4-[4-amino-2-(4-sulfamoyl-phenylamino)-thiazole-5-carbonyl]-benzoate(500 mg, 1.12 mmol; Example C(34)) in MeOH (10 mL) was added 1N aq NaOH(3.4 mL, 3.4 mmol). After 4 hours, the resultant mixture was acidifiedwith 1N aq HCl to pH 3 and filtered. The isolated brown solidcrystallized in EtOH to provide 330 mg (70% yield) of light browncrystals, mp 298.5-300° C.

[1174]¹H NMR (DMSO-d₆): δ 13.15 (1H, s), 11.14 (1H, s), 8.31 (2H, bs),8.02 (2H, d, J=8.1 Hz), 7.78 (4H, s), 7.77 (2H, d, J=8.1 Hz), 7.26 (2H,s).

[1175] HRFABMS (M+Na⁺): Calcd.: 441.0303. Found: 441.0320.

[1176] Anal. Calcd. for C₁₇H₁₄N₄O₅S₂.0.4H₂O: C, 47.97; H, 3.50; N,13.16; S, 15.07. Found: C, 48.04; H, 3.48; N, 12.98; S, 15.18.

Example F2-[4-Amino-2-(4-methoxy-phenylamino)-thiazole-5-carbonyl]-benzonitrile

[1177]

[1178] To a solution of the compound of Example C(12) (2.00 g, 4.43mmol) in pyridine (5 mL) was added copper(I) cyanide (709 mg, 8.86mmol), and the mixture was heated to reflux. After 2 hours, theresultant mixture was allowed to cool, acidified with 1N aqueous HCl,and extracted with 20% MeOH/CHCl₃. The CHCl₃ extracts were combined,washed with H₂O and brine, dried over Na₂SO₄, and evaporated to providea dark-brown viscous oil, which was purified via preparative thin-layerchromatography with 5% MeOH/CH₂Cl₂ and precipitated from EtOH to furnish255 mg (61% yield) of yellow amorphous solid that decomposed at 110-116°C.

[1179]¹H NMR (DMSO-d₆): δ 10.70 (1H, s), 8.24 (2H, bs), 7.91 (1H, d,J=7.8 Hz), 7.80-7.66 (2H, m), 7.61 (1H, td, J=7.8, 1.2 Hz), 7.42 (2H, d,J=9.0 Hz), 6.92 (2H, d, J=9.0 Hz), 3.72 (3H, s).

[1180] FABMS (MH⁺): 351.

[1181] Anal. Calcd. for C₁₈H₁₄N₄O₂S.0.25H₂O.0.2EtOH: C, 60.69; H, 4.35;N, 15.39; S, 8.81. Found: C, 60.84; H, 4.24; N, 15.07; S, 9.02.

Example G[4-Amino-2-(1H-benzoimidazol-6-ylamino)-thiazol-5-yl]-(3-amino-2,6-dichloro-phenyl)-methanone

[1182]

[1183] The title compound of Example C(82),N-{3-[4-amino-2-(1H-benzoimidazol-6-ylamino)-thiazole-5-carbonyl]-2,4-dichloro-phenyl}-acetamide(100 mg, 0.220 mmol), was placed in 6N aq. HCl (4 mL) and stirred atambient temperature for 24 hours. The mixture was brought to pH 7 with2N aq NaOH and the resultant pale yellow precipitate was filtered off,washed with H₂O, recrystallized from MeOH/H₂O, and dried under highvacuum. A yellow solid was obtained in 36% yield, mp 235-237° C.

[1184]¹H NMR (DMSO-d₆): δ 8.16 (1H, bs), 7.86 (2H, bs), 7.38-7.62 (1H,m), 7.18 (1H, d, J=8.5 Hz), 7.02 (1H, d, J=8.8 Hz), 6.68 (11H, d, J=8.7Hz), 5.50 (1H, bs).

[1185] IR (KBr): 3177, 1614, 1543, 1443, 1308 cm⁻¹.

[1186] FABMS (MH⁺): 419.

[1187] Anal. Calcd. for C₁₇H₁₂Cl₂N₆OS.0.8H₂O.1MeOH: C, 46.42; H, 3.81;N, 18.04; S, 6.88. Found: C, 46.37; H, 3.45; Cl, 15.29; N, 17.84; S,6.77.

Example H(1)[4-Amino-2-(4-piperazin-1-yl-phenylamino)-thiazol-5-yl]-(3-methyl-thiophen-2-yl)-methanoneTrihydrochloride

[1188]

[1189] The title compound was prepared as follows. To a solution of thetitle compound of Example C(104) (100 mg, 0.20 mmol) in a mixture of THF(1 mL) and MeOH (0.5 mL) was added a solution of 4N HCl in dioxane (200μL, 0.80 mmol). The resultant suspension was heated at reflux for 2hours. The suspension was allowed to cool and filtered. The isolatedsolid was washed with anhydrous ether and dried to provide a yellowsolid in 97% yield, mp 198-200° C.

[1190]¹H NMR (DMSO-d₆): δ 10.80 (1H, m), 9.22 (1H, bs), 7.60 (1H, d,J=5.0 Hz), 7.42 (1H, d, J=8.7 Hz), 6.98-7.08 (3H, m), 3.38 (4H, d, J=4.4Hz), 3.22 (4H, s), 2.18 (3H, s).

[1191] IR (KBr): 3177, 1614, 1543, 1443, 1308 cm⁻¹.

[1192] HRFABMS: Calcd. for C₁₉H₂₂N₅OS₂ (MH⁺): 400.1266. Found: 400.1254.

[1193] Anal. Calcd. for C₁₉H₂₁N₅OS₂.0.6H₂O.3HCl: C, 43.91; H, 4.89; N,13.47; S, 12.34. Found: C, 43.61; H, 4.97; N, 13.12; S, 12.16.

Example H(2)(3-Amino-2,6-dichloro-phenyl)-[4-amino-2-(4-piperazin-1-yl-phenylamino)-thiazol-5-yl]-methanoneTrihydrochloride

[1194]

[1195] The title compound was prepared in a manner like that describedfor Example H(1). The title compound of Example C(106) provided a yellowsolid in 48% yield, mp>280° C.

[1196]¹H NMR (DMSO-d₆): δ 8.88 (1H, bs), 8.00 (1H, bs), 7.40(2H, bs),7.18 (1H, d, J=8.7 Hz), 6.98 (2H, d, J=8.4 Hz), 6.80 (1H, d, J=8.7 Hz),3.38 (4H, s), 3.12 (4H, s).

[1197] IR (KBr): 3406, 1618, 1560, 1458, 1308 cm⁻¹.

[1198] HRFABMS: Calcd. for C₂₀H₂₁Cl₂N₆OS (MH⁺): 463.0875. Found:463.0862.

[1199] Anal. Calcd. for C₂₀H₂₀Cl₂N₆OS.3HCl.0.5 dioxane: C, 42.84; H,4.41; Cl, 28.74; N, 13.62; S. 5.20. Found: C, 42.96; H, 4.47; Cl, 28.58;N, 13.53; S. 5.15.

Example H(3)[4-Amino-2-(4-piperazin-1-yl-phenylamino)-thiazol-5-yl]-(2,6-dichloro-phenyl)-methanone

[1200]

[1201] The title compound was prepared in a manner like that describedfor Example H(1). The title compound of Example C(105) provided a yellowsolid in 44% yield, mp 298-300° C.

[1202]¹H NMR (DMSO-d₆): δ 7.60-7.50 (5H, m), 7.08 (2H, d, J=7.8 Hz),3.44 (4H, bs).

[1203] IR (KBr): 3395, 2959, 1618, 1513, 1425 cm⁻¹.

[1204] HRFABMS: Calcd. for C₂₀H₂₀Cl₂N₅OS (MH⁺): 448.0766. Found:448.0749.

[1205] Anal. Calcd. for C₂₀H₁₉Cl₂N₅OS.1.2H₂O.0 9HCl: C, 47.78; H, 4.47;Cl, 20.45; N, 13.93; S, 6.38. Found: C, 47.99; H, 4.38; Cl, 20.57; N,13.56; S, 6.24.

Example J(1)[4-Amino-2-(4-piperazin-1-yl-phenylamino)-thiazol-5-yl]-(2,4,6-trichloro-phenyl)-methanone

[1206]

[1207]{4-Amino-2-[4-(4-t-butoxycarbonyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(2,4,6-trichloro-phenyl)-methanone,which has the structural formula

[1208] was prepared essentially as described for Example C(1).1-t-Butoxycarbonyl-4-(4-isothiocyanato-phenyl)-piperazine (from ExampleC(101)) and 2-bromo-2′,4′,6′-trichloroacetophenone (from Example C(107))gave a black tar, which precipitated from EtOH to give 144 mg (50%) ofyellow amorphous powder, mp 192-193° C. (d).

[1209]¹H NMR (DMSO-d₆): δ 7.78 (2H, s), 7.33 (2H, bm), 6.98 (2H, d,J=9.0 Hz), 3.15-3.05 (4H, m), 1.45 (s, 9H).

[1210] IR (KBr): 3389, 3276, 3166, 1676, 1608, 1577, 1544, 1461, 1421,1366, 1235, 1202, 1164 cm⁻¹.

[1211] HRFABMS: Calcd for C₂₅H₂₆Cl₃N₅O₃SCs (M+Cs⁺): 715.9847. Found:715.9822.

[1212] Anal. Calcd for C₂₅H₂₆Cl₃N₅O₃S.0.75H₂O.0.4EtOH: C, 50.40; H,4.90; N, 11.39; Cl, 17.30; S, 5.22. Found: C, 50.69; H, 5.16; N, 10.98;Cl, 17.70; S, 4.90.

[1213] The title compound was prepared as follows.{4-Amino-2-[4-(4-tert-butoxycarbonyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(2,4,6-trichloro-phenyl)-methanone(50 mg, 0.086 mmol) was stirred in trifluoroacetic acid (TFA; 0.5 mL) at0° C. After 20 min at 0° C., a minimal amount of water was added, andsat aq NaHCO₃ was used for neutralization. The resultant suspension wasfiltered to obtain a yellow paste, which gave a suspension withMeOH/CHCl₃ and led to isolation of 22 mg (42%) of yellow amorphouspowder.

[1214]¹H NMR (DMSO-d₆): δ 7.80 (2H, s), 7.38 (2H, d, J=9.0 Hz), 7.01(2H, d, J=9.0 Hz).

[1215] IR (KBr): 3396, 3284, 3178, 1676, 1614, 1543, 1461, 1423, 1202,1137 cm⁻¹.

[1216] HRFABMS: Calcd for C₂₀H₁₈Cl₃N₅OS (MH⁺): 484.0346. Found:484.0333.

[1217] Anal. Calcd for C₂₀H₁₈Cl₃N₅OS.0.8MeOH.0.8 CHCl₃: C, 42.96; H,3.67; N, 11.60. Found: C, 42.87; H, 3.45; N, 11.27.

Example J(2)[4-Amino-2-(4-piperazin-1-yl-phenylamino)-thiazol-5-yl]-(2,6-difluoro-phenyl)-methanone

[1218]

[1219] The title compound was prepared essentially as described forExample J(1). To the title compound of Example C(101) (250 mg, 0.48mmol) in CH₂Cl₂ at 0° C. was added TFA (5 mL). After 20 min at 0° C.,the resultant clear solution was concentrated in vacuo to a residuewhich was suspended in a minimal amount of water, cooled to 0° C., andbasified with sat. Na₂CO₃ to pH 9. The solid was collected andrecrystallized from EtOH to obtain 116 mg (58% yield) of yellow solid,mp 190-193° C.

[1220]¹H NMR (DMSO-d₆): δ 8.13 (2H, bs), 7.52 (1H, p, J=7.3 Hz), 7.36(2H, d, J=8.7 Hz), 7.19 (2H, t, J=8.7 Hz), 6.99 (2H, t, d=8.7 Hz), 3.24(4H, bs), 3.13 (4H, bs).

[1221] HRFABMS (MH⁺): Calcd.: 416.1357. Found: 416.1370.

[1222] Anal. Calcd. for C₂₀H₁₉N₅OSF₂.0.7H₂O.0.7CF₃COOH: C, 49.96; H,4.11; N, 13.49; S, 6.17. Found: C, 50.16; H, 4.33; N, 13.14; S, 6.06.

Example J(3)[4-Amino-2-(4-piperazin-1-yl-phenylamino)-thiazol-5-yl]-(2,4,6-trifluoro-phenyl)-methanone

[1223]

[1224]{4-Amino-2-[4-(4-t-butoxycarbonyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(2,4,6-trifluoro-phenyl)-methanone,which has the structural formula

[1225] was prepared essentially as described for Example C(1).1-tert-Butoxycarbonyl-4-(4-isothio-cyanato-phenyl)-piperazine (fromExample C(101)) and 2′-bromo-2,4,6-trifluoroacetophenone (from ExampleC(115)) gave a yellow solid, which crystallized from EtOH to give 200 mg(80%) of yellow amorphous powder that darkened at 125-130° C., mp132-135° C. (decomposed).

[1226]¹H NMR (CD₃CN): δ 8.69 (1H, bs), 7.46 (2H, d, J=9.0 Hz), 7.20-7.10(4H, m), 3.74-3.62 (4H, m), 3.28-3.20 (4H, m), 1.60 (s, 9H).

[1227] IR(KBr): 3389, 3282, 3178, 1686, 1637, 1604, 1546, 1427, 1366,1343, 1233, 1168, 1121, 1035, 999 cm⁻¹.

[1228] HRFABMS: Calcd for C₂₅H₂₇F₃N₅O₃S (MH⁺): 534.1787. Found:534.1772.

[1229] Anal. Calcd for C₂₅H₂₆F₃N₅O₃S.1H₂O.0.5EtOH: C, 54.35; H, 5.44; N,12.19; S, 5.58. Found: C, 54.26; H, 5.07; N, 11.92; S, 5.50.

[1230] The title compound was prepared essentially as described forExample J(1) to give a brown solid, which was purified via columnchromatography with 10% MeOH/CHCl₃ as eluant to provide 57 mg (60%) of ayellow-orange amorphous solid that decomposed above 205° C.

[1231]¹H NMR (CD₃CN): δ 7.78 (2H, s), 7.42 (2H, d, J=9.0 Hz), 7.01 (2H,d, J=9.0 Hz), 3.30-3.18 (4H, m), 3.14-3.02 (4H, m).

[1232] IR(KBr): 33406, 1603, 1544, 1430, 1237, 1120, 1034 cm¹.

[1233] HRFABMS: Calcd for C₂₀H₁₈F₃N₅OS (MH⁺): 434.1262. Found: 434.1274.

[1234] Anal. Calcd for C₂₀H₁₈F₃N₅OS.0.7MeOH.0.7CHCl₃: C, 47.65; H, 4.02;N, 12.98; S, 5.94. Found: C, 47.84; H, 3.64; N, 12.59; S, 5.69.

Example J(4)4-[4-Amino-5-(2,6-difluoro-benzoyl)-thiazol-2-ylamino]-N-piperidin-4-ylmethyl-benzenesulfonamide

[1235]

[1236] N-tert-Butoxycarbonyl-4-carbamoyl-piperidine, which has thestructural formula

[1237] was made as follows. To isonipecotamide (5.00 g, 39.0 mmol) indioxane (100 mL) was added di-tert-butyl dicarbonate (8.51 g, 39.0 mmol)and N,N-diisopropylethylamine (6.0 mL, 42.9 mmol). The mixture wasallowed to stir overnight, then evaporated undr reduced pressure todryness. The residue was partitioned between CHCl₃ and 1N HCl. Theorganic layer was washed with water and brine, dried over Na₂SO₄, andconcentrated to give 8.3 g (93% yield) of white solid, which was usedwithout further purification.

[1238]¹H NMR (CDCl₃): δ 5.53 (2H, bs), 4.03 (2H, d, J=13.7 Hz), 2.33(2H, tt, J=11.8, 3.7 Hz), 2.08 (2H, bs), 1.89 (2H, dd, J=13.7, 3.7 Hz),1.69 (1H, dd, J=11.8, 4.4 Hz), 1.65-1.57 (1H, m), 1.44 (9H, s).

[1239] 4-Aminomethyl-N-tert-butoxycarbonyl-piperidine, which has thestructural formula

[1240] was made as follows. ToN-tert-butoxycarbonyl-4-carbamoyl-piperidine (15.6 mmol) in THF (40 mL)at −78° C. under Ar was added LiAlH₄ (592 mg, 15.6 mmol). The mixturewas allowed to warm to ambient temperature slowly and after a half hour,recooled to −78° C., quenched with ethyl acetate, and partitionedbetween EtOAc and 2N NaOH. The organic layer was separated, dried overK₂CO₃, and concentrated to give 1.98 g (59% yield) of yellow slurry,which was used without further purification.

[1241]N-tert-Butoxycarbonyl-4-[(4-nitro-benzenesulfonylamino)-methyl]-piperidine,which has the structural formula

[1242] was made as follows. 4-nitrobenzenesulfonyl chloride (2.05 g,9.24 mmol) was added to a solution of4-aminomethyl-N-tert-butoxycarbonyl-piperidine (1.98 g, 9.24 mmol) inTHF (20 mL) at ambient temperature. The mixture was refluxed for 1 hour,concentrated in vacuo, and partitioned between CH₂Cl₂ and 1N HCl. Theorganic layer was washed with brine, dried over Na₂SO₄, passed through apad of silica gel, and concentrated to give 1.71 g (46% yield) of yellowsolid, which was used without further purification.

[1243]4-[(4-Amino-benzenesulfonylamino)-methyl]-N-tert-butoxycarbonyl-piperidine,which has the structural formula

[1244] was prepared as follows.N-tert-Butoxycarbonyl-4-[(4-nitro-benzenesulfonylamino)-methyl]-piperidine(1.70 g, 4.26 mmol), 10% Pd/C (250 mg), MeOH (10 mL), and THF (10 mL)was stirred under hydrogen for 2 hours and filtered. The filtrate wasconcentrated to a residue that was purified via column chromatographywith 5% MeOH/CHCl₃ as eluant, producing 1.39 g (88% yield) of whitesolid, which was used without further purification.

[1245]N-tert-Butoxycarbonyl-4-[(4-isothiocyanato-benzenesulfonylamino)-methyl]-piperidine,which has the structural formula

[1246] was prepared in a manner analogous to1-(4-isothiocyanato-phenyl)-morpholine for Example C(54).4-[(4-Amino-benzenesulfonylamino)-methyl]-N-tert-butoxycarbonyl-piperidineprovided a yellow solid in 39% yield, which was used without furtherpurification.

[1247]4-{[4-(5-Acetyl-4-amino-thiazol-2-ylamino)-benzenesulfonylamino]-methyl}-N-tert-butoxycarbonyl-piperidine,which has the structural formula

[1248] was prepared in a manner analogous to that used in Example C(1).N-tert-Butoxycarbonyl-4-[(4-isothiocyanato-benzenesulfonylamino)-methyl]-piperidineand 2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided ayellow solid in 50% yield.

[1249]¹H NMR (DMSO-d₆): δ 11.22 (1H, s), 8.20 (2H, bs), 7.84-7.73 (3H,m), 7.62-7.54 (2H, m), 7.24 (2H, dd, J=7.8, 7.7 Hz), 3.89 (2H, d, J=12.8Hz), 3.35 (2H, s), 2.52 (2H, d, J=1.2 Hz), 1.60 (2H, d, J=10.1 Hz),1.56-1.42 (1H, m), 1.39 (9H, s), 0.91 (2H, d, J=12.8 Hz).

[1250] The title compound was prepared in a manner analogous to thatused in Example J(1).4-{[4-(5-Acetyl-4-amino-thiazol-2-ylamino)-benzenesulfonylamino]-methyl}-N-tert-butoxycarbonyl-piperidineprovided a brown solid in 28% yield.

[1251]¹H NMR (DMSO-d₆): δ 8.11 (2H, bs), 7.70 (4H, bs), 7.58-7.42 (1H,m), 7.20 (1H, d, J=7.8 Hz), 7.15 (1H, d, J=7.8 Hz), 3.80 (2H, bs), 3.05(2H, d, J=10.0 Hz), 2.60 (2H, d, J=6.8 Hz), 1.65 (2H, d, J=12.2 Hz),1.52 (1H, bs), 1.07 (2H, d, J=10.0 Hz).

[1252] HRFABMS (MH⁺): Calcd.: 507.1210. Found: 507.1206.

[1253] Anal. Calcd. for C₂₂H₂₃N₅O₃S₂F₂.0.1CH₃OH.0.2CF₃COOH: C, 50.65; H,4.73; N, 13.12; S, 12.02. Found: C, 50.92; H, 4.46; N, 12.87; S, 12.18.

Example J(5){4-Amino-2-[4-(cis-3,5-dimethyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(2,6-difluoro-phenyl)-methanone

[1254]

[1255] 2,6-cis-dimethyl-4-(4-nitro-phenyl)-piperazine, which has thestructural formula

[1256] was first prepared essentially as described for3R-methyl-1-(4-nitro-phenyl)-piperazine for Example C(124).cis-2,6-Dimethylpiperazine gave 2.19 g (100% yield) of yellow powder mp130-131.5° C., which was used without further purification.

[1257]¹H NMR (CDCl₃): δ 8.03 (2H, d, J=9.5 Hz), 7.02 (2H, d, J=9.5 Hz),3.88 (2H, dd, J=12.4, 2.0 Hz), 2.82-2.68 (2H, m), 2.44-2.33 (3H, m),1.03 (6H, d, J=6.3 Hz).

[1258] IR(KBr): 1596, 1509, 1482, 1316, 1252, 1193, 1119, 1101 cm¹.

[1259] Anal. Calcd. for C₁₂H₁₇N₃O₂: C, 61.26; H, 7.28; N, 17.86. Found:C, 61.25; H, 7.42, N, 17.84.

[1260] 1-tert-Butoxycarbonyl-2,6-dimethyl-4-(4-nitro-phenyl)-piperazine,which has the structural formula

[1261] was prepared as follows. To2,6-cis-dimethyl-4-(4-nitro-phenyl)-piperazine (1.00 g, 4.25 mmol) indioxane (20 mL) was added di-tert-butyl dicarbonate (1.12 g, 5.12 mmol)and N,N-diisopropylethylamine (1.37 mL, 9.76 mmol). After 3 hours at 80°C., the mixture was allowed to cool and evaporated to dryness. The solidwas suspended in water, filtered off, washed with water, and dried undervacuum to give 1.40 g (98% yield) a yellow powder, which was usedwithout further purification.

[1262]¹H NMR (CDCl₃): δ 8.12 (2H, ddd, J=7.3, 2.1, 2.1 Hz), 6.80 (2H,ddd, J=7.3, 2.1, 2.1 Hz), 4.30 (2H, ddd, J=13.2, 6.8, 4.5 Hz), 3.71 (d,2H, J=13.2 Hz), 3.22 (dd, 2H, J=12.8, 4.5 Hz), 1.49 (9H, s), 1.29 (6H,d, J=6.8 Hz).

[1263] IR (KBr): 1689, 1594, 1489, 1400, 1322, 1257, 1057 cm⁻¹.

[1264] 1-(4-Amino-phenyl)-4-tert-butoxycarbonyl-3,5-dimethyl-piperazine,which has the structural formula

[1265] was prepared as follows. Hydrogenation of crude1-tert-butoxycarbonyl-2,6-dimethyl-4-(4-nitro-phenyl)-piperazine (1.48g, 4.41 mmol) in THF (20 mL) and MeOH (20 mL) with 10% Pd/C as catalystgave 1.12 g (83% yield) of a clear sticky oil, which was used withoutfurther purification.

[1266]¹H NMR (CDCl₃): δ 8.13 (2H, d, J=9.4 Hz), 6.81 (2H, d, J=9.4 Hz),4.30 (2H, ddd, J=13.2, 6.8, 4.5 Hz), 3.71 (2H, d, J=13.2 Hz), 3.21 (2H,dd, J=13.2, 4.5 Hz), 1.49 (9H, s), 1.29 (6H, d, J=6.8 Hz).

[1267]1-(tert-Butoxycarbonyl)-2,6-cis-dimethyl-4-(4-isothiocyanato-phenyl)-piperazine,which has the structural formula

[1268] was prepared in a manner analogous to4-(4-isothiocyanatophenyl)-morpholine for Example C(54).1-(4-Amino-phenyl)-4-tert-butoxycarbonyl-3,5-dimethyl-piperazineprovided a clear sticky foam that recrystallized from cold ether/hexanesto furnish pale tan crystals in 68% yield, mp 97-98° C.

[1269]¹H NMR (CDCl₃): δ 6.74 (2H, d, J=8.7 Hz), 6.67 (2H, d, J=8.7 Hz),4.20-4.08 (2H, m), 3.08 (2H, d, J=11.6 Hz), 2.71 (2H, dd, J=11.6, 3.9Hz), 1.41 (9H, s), 1.28 (6H, d, J=6.8 Hz).

[1270] IR(KBr): 2175, 2135, 1691, 1507, 1395, 1341, 1246, 1177, 1098cm⁻¹.

[1271] Anal. calcd for C₁₈H₂₅N₃O₂S: C, 62.21; H, 7.25; N, 12.09; S,9.23. Found: C, 62.31; H, 7.32; N, 11.96; S, 9.39.

[1272]4-Amino-2-[4-(1-tert-butoxycarbonyl-2,6-cis-dimethyl-piperazine-4-yl)-phenylamino]-thiazol-5-yl-(2,6-difluorophenyl)-methanone,which has the structural formula

[1273] was prepared in a manner analogous to that used in Example C(1).1-(tert-Butoxycarbonyl)-2,6-cis-dimethyl-4-(4-isothiocyanato-phenyl)-piperazineand 2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided ayellow solid in 51% yield, which was used without further purification.

[1274]¹H NMR (DMSO-d₆): δ 10.66 (1H, s), 8.12 (2H, bs), 7.56-7.44 (1H,m), 7.38 (2H, d, J=9.0 Hz), 7.18 (1H, d, J=7.7 Hz), 7.15 (1H, d, J=8.1Hz), 6.95 (2H, d, J=9.0 Hz), 4.14-4.03 (2H, m), 3.49-3.41 (2H, m), 2.75(2H, dd, J=12.2, 4.4 Hz), 1.42 (9H, S), 1.24 (6H, d, J=6.7 Hz).

[1275] FABMS (M+Na⁺): 566

[1276] The title compound was prepared in a manner analogous to thatused in Example J(1).4-Amino-2-[4-(1-tert-butoxycarbonyl-2,6-dimethyl-piperazine-4-yl)-phenylamino]-thiazol-5-yl-(2,6-difluorophenyl)-methanoneprovided a brown powder in 52% yield, mp 293-294.5° C.

[1277]¹H NMR (DMSO-d₆): δ 8.11 (2H, bs), 7.56-7.44 (1H, m), 7.26 (2H, d,J=9.0 Hz), 7.18 (1H, d, J=7.7 Hz), 7.14 (1H, d, J=8.1 Hz), 6.89 (2H, d,J=9.0 Hz), 3.48 (2H, dd, J=10.9, 2.2 Hz), 2.88-2.76 (2H, m), 2.07 (4H,t, J=10.9 Hz), 1.00 (6H, d, J=6.3 Hz).

[1278] HRFABMS (MH⁺): Calcd.: 444.1670. Found: 444.1658.

[1279] Anal. Calcd. for C₂₂H₂₃N₅OSF₂.0.4H₂O: C, 58.63; H, 5.32; N,15.54; S, 7.11. Found: C, 58.64; H, 5.40; N, 15.23; S, 6.96.

Example J(6){4-Amino-2-[4-(3,3-dimethyl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(2,6-difluoro-phenyl)-methanone

[1280]

[1281] 2,2-Dimethyl-4-(4-nitro-phenyl)-piperazine, which has thestructural formula

[1282] was first prepared as follows. Crude 2,2-dimethylpiperazine (10.0mmol; Chu et al., Can. J. Chem., vol. 70 (1992), pp. 1328-1337),4-fluoro-nitrobenzene (5.0 mmol, 706 mg), and K₂CO₃ (8.3 g, 60.0 mmol)in DMSO (10 mL) was heated at 100° C. for 4 hours, cooled, diluted withwater (100 mL), and extracted with ether:ethyl acetate (200:50 mL). Theorganic layer was washed with water (3×) and brine, and concentrated toprovide 1.17 g (100% yield) of yellow solid, which was used withoutfurther purification.

[1283]¹H NMR (CDCl₃): δ 8.13 (2H, d, J=9.5 Hz), 6.80 (2H, d, J=9.5 Hz),3.38 (2H, dd, J=5.5, 5.0 Hz), 3.20 (2H, s), 3.07 (2H, dd, J=5.5, 5.0Hz), 1.21 (6H, s).

[1284] 1-tert-Butoxycarbonyl-2,2-dimethyl-4-(4-nitro-phenyl)-piperazine,which has the structural formula

[1285] was prepared in a manner analogous to1-(4-amino-phenyl)-4-(tert-butoxycarbonyl)-2,6-dimethyl-piperazine forExample J(5). 2,2-Dimethyl-4-(4-nitro-phenyl)-piperazine provided abright yellow solid in 99% yield, which was used without furtherpurification.

[1286]¹H NMR (CDCl₃): δ 8.15 (2H, d, J=9.4 Hz), 6.64 (2H, d, J=9.4 Hz),3.90 (2H, dd, J=6.0, 5.5 Hz), 3.54 (2H, dd, J=6.0, 5.5 Hz), 3.53 (2H,s), 1.51 (9H, s), 1.44 (6H, s).

[1287]1-(4-Amino-phenyl)-4-(tert-butoxycarbonyl)-3,3-dimethyl-piperazine,which has the structural formula

[1288] was prepared as follows.1-tert-Butoxycarbonyl-2,2-dimethyl-4-(4-nitro-phenyl)-piperazine (700mg, 2.09 mmol) and 10% Pd/C (100 mg) in THF (15 mL) and MeOH (15 mL) wasstirred under hydrogen for 2 hours and filtered. The filtrate wasconcentrated in vacuo to give a light brown slurry, which was usedwithout further purification.

[1289]¹H NMR (CDCl₃): δ 6.69-5.65 (4H, m), 3.67 (2H, dd, J=5.8, 5.4 Hz),3.21-3.14 (2H, m), 3.01 (2H, s), 1.49 (9H, s), 1.43 (6H, s).

[1290]1-(tert-Butoxycarbonyl)-2,2-dimethyl-4-(4-isothiocyanato-phenyl)-piperazine,which has the structural formula

[1291] was prepared analogous to 4-isothiocyanato-benzamide for C(102).1-(4-Amino-phenyl)-4-(tert-butoxycarbonyl)-3,3-dimethyl-piperazineprovided a white solid in 80% yield, which was used without furtherpurification.

[1292]¹H NMR (CDCl₃): δ 7.15 (2H, d, J=9.0 Hz), 6.63 (2H, d, J=9.0 Hz),3.85 (2H, dd, J=5.9, 5.5 Hz), 3.42 (2H, dd, J=5.9, 5.5 Hz), 3.37 (2H,s), 1.57 (9H, s), 1.44 (6H, s).

[1293]4-Amino-2-[4-(1-tert-butoxycarbonyl-2,2-dimethyl-piperazine-4-yl)-phenylamino]-thiazol-5-yl-(2,6-difluorophenyl)-methanone,which has the structural formula

[1294] was prepared in a manner analogous to that used in Example C(1).1-(tert-Butoxycarbonyl)-2,2-dimethyl-4-(4-isothiocyanato-phenyl)-piperazineand 2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided ayellow powder in 60% yield, which was used without further purification.

[1295]¹H NMR (DMSO-d₆): δ 10.58 (1H, s), 8.13 (2H, bs), 7.61-7.48 (1H,m), 7.40-7.15 (5H, m), 6.79 (2H, d, J=9.1 Hz), 3.74 (2H, dd, J=5.8, 5.3Hz), 3.41-3.30 (4H, m), 1.48 (9H, s), 1.39 (6H, s).

[1296] The title compound was prepared in a manner analogous to thatused in Example J(1).4-Amino-2-[4-(1-tert-butoxycarbonyl-2,2-dimethyl-piperazine-4-yl)-phenylamino]-thiazol-5-yl-(2,6-difluorophenyl)-methanoneprovided a yellow solid in 51% yield, mp 205-210° C.

[1297]¹H NMR (DMSO-d₆): δ 8.15 (2H, bs), 7.63-7.54 (1H, m), 7.35 (2H, d,J=9.0 Hz), 7.25 (1H, d, J=7.7 Hz), 7.22 (1H, d, J=8.1 Hz), 6.98 (2H, d,J=9.0 Hz), 3.10-3.04 (2H, m), 3.02-2.95 (2H, m), 2.92 (2H, s), 1.21 (6H,s).

[1298] IR (KBr): 3276, 2961, 1620, 1590, 1546. 1516, 1464, 1429, 1364,1257, 1232, 1002 cm⁻¹.

[1299] HRFABMS (MH⁺): Calcd.: 444.1670. Found: 444.1657.

[1300] Anal. Calcd. for C₂₂H₂₃N₅OSF₂.0.7CH₃OH: C, 58.51; H. 5.58; N,15.03; S. 6.88. Found: C, 58.601; H, 5.68; N, 14.87; S, 6.76.

Example K{4-Amino-2-[4-(4-pyridin-2-yl-piperazin-1-yl)-phenylamino]-thiazol-5-yl}-(2,6-difluoro-phenyl)-methanone

[1301]

[1302] 1-(4-Nitro-phenyl)-4-pyridin-2-yl-piperazine, which has thestructural formula

[1303] was first prepared in a manner analogous to tert-butyl[methyl-(4-nitro-phenyl)-amino]-acetate for Example C(103).1-Pyridin-2-yl-piperazine and 4-fluoronitrobenzene gave a yellow solidin 85% yield.

[1304]¹H NMR (CDCl₃): δ 8.13-8.28 (3H, m), 7.50-7.58 (2H, m), 7.52 (1H,ddd, J=15.7, 7.3, 2.0 Hz), 6.88 (2H, d, J=9.4 Hz), 6.70 (2H, dd, J=7.4,5.1 Hz), 3.78 (4H, dd, J=7.4, 5.0 Hz), 3.62 (4H, dd, J=5.7, 3.3 Hz).

[1305] 4-(1-Pyridin-2-yl-piperazin-4-yl)-aniline, which has thestructural formula

[1306] was prepared in a manner analogous to4-(4-methyl-piperazin-1-yl)-aniline for Example C(70).1-(4-Nitro-phenyl)-4-pyridin-2-yl-piperazine afforded a gray solid in94% crude yield, which was used without further purification.

[1307]¹H NMR (CDCl₃): δ 8.22 (1H, bd, J=3.5 Hz), 7.52 (1H, ddd, J=17.6,7.2, 1.9 Hz), 6.88 (2H, d, J=8.7 Hz), 6.62-6.78 (4H, m), 3.72 (4H, dd,J=5.2, 5.0 Hz), 3.48 (2H, bs), 3.18 (4H, t, J=5.2, 5.0 Hz).

[1308] 1-(4-Isothiocyanato-phenyl)-4-pyridin-2-yl-piperazine, which hasthe structural formula

[1309] was prepared in a manner analogous to1-(4-isothiocyanato-phenyl)-4-pyridin-4-yl-piperazine for ExampleC(127). 4-(4-Pyridin-2-yl-piperazin-1-yl)-aniline gave 2.2 g (95% yield)of a yellow solid, which was used without any further purification.

[1310]¹H NMR (CDCl₃): δ 8.26 (1H, bd, J=6.3 Hz), 7.91 (1H, ddd, J=18.1,7.1, 1.8 Hz), 7.18 (2H, d, J=9.0 Hz), 6.82-7.00 (4H, m), 4.10 (4H, dd,J=5.3, 5.1 Hz), 3.48 (4H, dd, J=5.3, 5.2 Hz).

[1311] The title compound was prepared as follows. To a solution of1-(4-isothiocyanato-phenyl)-4-pyridin-2-yl-piperazine (250 mg, 0.84mmol) in dry MeOH (4 mL) was added cyanamide (35 mg, 0.84 mmol) and afresh solution of NaOH (67 mg, 1.67 mmol) in dry MeOH (4 mL). After 1hour, 2-bromo-2′,6′-difluoro-acetophenone (from Example C(79); 178 mg,0.76 mmol) was added. The next day, the resultant yellow suspension wasfiltered. The solid was washed with H₂O and dried under high vacuum toafford a yellow solid in 86% yield, mp 138-140° C.

[1312]¹H NMR (DMSO-d₆): δ 8.12 (2H, dd, J=6.5, 1.7 Hz), 7.42-7.60 (2H,m), 7.32 (2H, bd, J=8.5 Hz), 7.08 (2H, t, J=9.0 Hz), 6.98 (2H, d, J=9.0Hz), 6.88 (2H, d, J=8.7 Hz), 6.64 (1H, dd, J=7.0, 5.0 Hz), 3.62 (4H, t,J=4.7 Hz), 3.20 (4H, t, J=4.7 Hz).

[1313] IR (KBr): 3369, 3180, 2835, 1620, 1597, 1546, 1466, 1433, 1232cm⁻¹.

[1314] HRFABMS: Calcd. for C₂₅H₂₃F₂N₆OS (MH⁺): 493.1622. Found:493.1608.

[1315] Anal. Calcd. for C₂₅H₂₂F₂N₆OS.0.9H₂O: C, 58.90; H, 4.90; N,16.49; S, 6.29. Found: C, 58.91; H, 4.64; N, 16.55; S, 6.24.

Example L{4-Amino-2-[4-(4-carboxamido-piperidin-1-yl)-phenylamino]-thiazol-5-yl}-(2,6-difluoro-phenyl)-methanone

[1316]

[1317] 4-Carboxamido-1-(4-nitro-phenyl)-piperidine, which has thestructural formula

[1318] was prepared in a manner analogous to tert-butyl[methyl-(4-nitro-phenyl)-amino]-acetate for Example C(103).4-Fluoronitrobenzene and isonipecotamide gave a yellow powder in 98%crude yield, which was used without further purification.

[1319]¹H NMR (CD₃OD): δ 8.22 (2H, d, J=9.5 Hz), 7.12 (2H, d, J=9.5 Hz),4.20 (2H, d, J=12.5 Hz), 3.16 (2H, ddd, J=25.6, 13.3, 2.7 Hz), 2.62-2.70(1H, m), 2.02 (2H, bd, J=10.3 Hz), 1.85-1.95 (2H, m).

[1320] 1-(4-Amino-phenyl)-4-carboxamido-piperidine, which has thestructural formula

[1321] was prepared in a manner analogous to4-(4-methyl-piperazin-1-yl)-aniline for Example C(70).4-Carboxamido-1-(4-nitro-phenyl)-piperidine gave a pale yellow powder in100% crude yield, which was used without further purification.

[1322]¹H NMR (CD₃OD): δ 6.60 (2H, bs), 6.42 (2H, bs), 3.22 (2H, bs),2.38 (2H, bs), 2.02 (1H, bs), 1.72-1.92 (4H, m).

[1323] 4-Carboxamido-1-(4-isothiocyanato-phenyl)-piperidine, which hasthe structural formula

[1324] was prepared in a manner analogous to1-(4-isothiocyanato-phenyl)-4-pyridin-2-yl-piperazine for Example K(1).1-(4-Amino-phenyl)-4-carboxamido-piperidine to give a cream-coloredpowder in 93% yield, which was used without further purification.

[1325]¹H NMR (CDCl₃): δ 7.14 (2H, d, J=9.0 Hz), 6.86 (2H, d, J=9.0 Hz),5.50 (1H, bs), 5.30 (1H, bs), 3.74 (2H, d, J=12.8 Hz), 2.82 (2H, ddd,J=24.3, 12.5, 2.8 Hz), 2.30-2.40 (1H, m), 1.80-2.08 (4H, m).

[1326] The title compound was prepared as follows. To a solution of4-carboxamido-1-(4-isothiocyanato-phenyl)-piperidine (198 mg, 0.76 mmol)in MeOH (3 mL) was added cyanamide (32 mg, 0.76 mmol) and a solution ofsodium methoxide in MeOH (1.65 mL of 0.5 N, 0.83 mmol). After 30 min,2-bromo-2′,6′-difluoro-acetophenone (162 mg, 0.69 mmol; from ExampleC(79)) was added. After 2 hours, H₂O was added. The yellow precipitatewas filtered off, washed with water, and recrystallized from boilingMeOH to give 200 mg (63% in yield) of an amorphous yellow powder,mp>300° C.

[1327]¹H NMR (DMSO-d₆): δ 7.46-7.58 (1H, m), 7.28 (2H, dd, J=8.8, 7.5Hz), 7.16 (3H, dd, J=8.0, 7.7 Hz), 6.82 (2H, d, J=9.1 Hz), 3.68 (2H, bd,J=12.6 Hz), 3.64 (2H, ddd, J=23.7, 12.1, 2.8 Hz), 2.04-2.18 (1H, m),1.52-1.82 (4H, m).

[1328] HRFABMS: Calcd. for. C₂₂H₂₁F₂N₅O₂SNa (M+Na⁺): 480.1282. Found:480.1266.

[1329] Anal. Calcd. for C₂₂H₂₁F₂N₅O₂S.0.2H₂O: C, 57.31; H, 4.68; N,15.19; S, 6.95. Found: C, 57.25; H, 4.63; N, 15.31; S, 7.01.

Example M1-{4-[4-Amino-5-(2,6-difluoro-benzoyl)-thiazol-2-ylamino]-phenyl}-piperidine-4-carboxylicAcid

[1330]

[1331] 1-(4-Nitrophenyl)-piperidine-4-carboxylic acid, which has thestructural formula

[1332] was prepared in a manner analogous to tert-butyl[methyl-(4-nitro-phenyl)-amino]-acetate for Example C(103).4-Fluoronitrobenzene and isonipecotic acid. afforded a yellow powder in89% crude yield, which was used without further purification.

[1333]¹H NMR (CDCl₃): δ 8.00 (2H, d, J=10.8 Hz), 6.71 (2H, d, J=10.7Hz), 3.80 (1H, t, J=3.9 Hz), 3.72 (1H, t, J=3.8 Hz), 2.98 (2H, ddd, J24.3, 11.1, 3.0 Hz), 2.48-2.60 (1H, m), 1.88-2.02 (2H, m), 1.68-1.82(2H, m).

[1334] Benzyl 1-(4-nitrophenyl)-piperidine-4-carboxylate, which has thestructural formula

[1335] was prepared as follows. To a suspension of1-(4-nitro-phenyl)-piperidine-4-carboxylic acid (500 mg, 2.01 mmol) inacetonitrile (10 mL) was added K₂CO₃ (612 mg, 4.44 mmol) and benzylbromide (265 μL, 2.22 mmol). The resultant mixture was heated at refluxfor 2 hours, allowed to cool, and diluted with H₂O. The aqueous layerwas extracted with ether (2×50 mL). The combined organic layers weredried over MgSO₄, filtered, and concentrated in vacuo to give 470 mg(64% in crude yield) of a yellow solid, which was used without furtherpurification.

[1336]¹H NMR (CDCl₃): δ 8.13 (2H, d, J=9.4 Hz), 7.30-7.42 (5H, m), 6.83(2H, d, J=9.4 Hz), 5.18 (2H, s), 3.92 (2H, dd, J=3.9, 3.5 Hz), 3.10 (2H,ddd, J=24.5, 13.7, 2.9 Hz), 2.62-2.70 (1H, m), 2.08 (2H, dd, J=13.5, 3.5Hz), 1.84-1.94 (2H, m).

[1337] Benzyl 1-(4-aminophenyl)-piperidine-4-carboxylate, which has thestructural formula

[1338] was prepared as follows. To a solution of benzyl1-(4-nitro-phenyl)-piperidine-4-carboxylate (400 mg, 1.18 mmol) indioxane (5 mL) and ethanol (1 mL) was added tin(II) chloride dihydrate(1.06 g, 4.70 mmol). The resultant solution was heated at reflux for 4hours, allowed to cool, and to aggregate solids, a small amount ofCelite added. The mixture was brought to pH 8 with saturated aq NaHCO₃and filtered. The filtrate was diluted with H₂O (50 mL) and extractedwith 5% MeOH in CHCl₃ (2×50 mL). The combined organic layers were driedover MgSO₄, filtered, and concentrated in vacuo to furnish 400 mg (100%crude yield) of a cream-colored powder, which was used without furtherpurification.

[1339]¹H NMR (CDCl₃): δ 7.30 (5H, bs), 6.58 (2H, d, J=8.8 Hz), 6.42 (2H,d, J=8.8 Hz), 4.94 (2H, s), 3.28 (1H, dd, J=3.6, 3.1 Hz), 3.18 (1H, dd,J=3.6, 3.0 Hz), 2.46 (2H, ddd, J=23.2, 11.8, 2.8 Hz), 2.14-2.28 (1H, m),1.60-1.88 (4H, m).

[1340] Benzyl 1-(4-isothiocyanatophenyl)-piperidine-4-carboxylate, whichhas the structural formula

[1341] was prepared as follows. To a solution of benzyl1-(4-amino-phenyl)-piperidine-4-carboxylate (400 mg, 1.29 mmol) in THF(5 mL) at −35° C. was added in succession Et₃N (435 μL, 3, 12 mmol) andthiophosgene (108 μL, 1.42 mmol). The resultant mixture was allowed towarm to ambient temperature, stirred for 0.5 hour, diluted with H₂O (50mL), and extracted with CHCl₃ (2×50 mL). The combined organic layerswere dried over MgSO₄, filtered, and concentrated under reduced pressureto give 400 mg (92% in yield) of a yellow powder, which was used withoutfurther purification.

[1342]¹H NMR (CD₃OD): δ 8.10 (2H, d, J=9.5 Hz), 7.38 (5H, d, J=4.5 Hz),6.92 (2H, d, J=9.5 Hz), 5.18 (2H, s), 4.00 (1H, t, J=3.4 Hz), 3.96 (1H,dd, J=3.5, 3.2 Hz), 3.13 (2H, ddd, J=24.9, 13.8, 2.9 Hz), 2.71-2.77 (1H,m), 2.05 (2H, dd, J=14.1, 3.4 Hz), 1.74-1.83 (2H, m).

[1343] Benzyl1-{4-[4-amino-5-(2,6-difluorobenzoyl)-thiazol-2-ylamino]-phenyl}-piperidine-4-carboxylate,which has the structural formula

[1344] was prepared as prepared in a manner like that described for thetitle compound of Example C(1). Benzyl1-(4-isothiocyanato-phenyl)-piperidine-4-carboxylate and2-bromo-2′,6′-difluoro-acetophenone (from Example C(79)) provided brownpowder in 82% yield, and was used without further purification.

[1345]¹H NMR (DMSO-d₆): δ 7.30 (1H, m), 7.18 (2H, d, J=8.9 Hz), 6.92(2H, d, J=9.0 Hz), 4.96 (2H, s), 3.62 (2H, bd, J=9.2 Hz), 2.80 (2H, ddd,J=26.4, 14.1, 2.6 Hz), 2.36-2.58 (1H, m), 2.04 (2H, bd, J=3.1 Hz),1.80-1.92 (2H, m).

[1346] The title compound was prepared as follows. To a mixture ofbenzyl1-{4-[4-amino-5-(2,6-difluoro-benzoyl)-thiazol-2-ylamino]-phenyl}-piperidine-4-carboxylate(150 mg, 0.27 mmol) in ethanol (10 mL) was added 20% palladium(II)hydroxide on carbon (60 mg). The resultant mixture stirred under ahydrogen atmosphere for 48 hours. The catalyst was filtered onto a padof Celite and rinsed with ethanol. The filtrate was concentrated underreduced pressure, and minimal ethyl acetate and CHCl₃ were added toinduce precipitation. The solid was filtered off, washed with ethylacetate, and dried to give 40 mg (30%) of a pale blue amorphous powder,mp 275-277° C., which was used without further purification.

[1347]¹H NMR (DMSO-d₆): δ 8.10 (1H, bs), 7.46-7.58 (1H, m), 7.30 (2H,bd, J=7.5 Hz), 7.16 (2H, dd, J=8.0, 7.9 Hz), 6.92 (2H, d, J=9.1 Hz),3.58 (2H, bd, J=12.6 Hz), 2.52 (2H, dd, J=11.2, 10.4 Hz), 2.32-2.40 (1H,m), 1.88 (2H, bd, J=16.1 Hz), 1.58-1.70 (2H, m).

[1348] Anal. Calcd. for C₂₂H₂₀F₂N₄O₃S.0.9H₂O.0.1CHCl₃: C, 54.55; H,4.54; N, 11.51; S, 6.59. Found: C, 54.55; H, 4.30; N, 11.13; S, 6.40.

Example N(1)[4-Amino-2-(4-nitro-phenylamino)-thiazol-5-yl]-(2-hydroxy-phenyl)-methanone

[1349]

and Example N(2)N-[5-(2-Hydroxy-benzoyl)-2-(4-nitro-phenylamino)-thiazol-4-yl]-benzamide

[1350]

[1351] Both title compounds were obtained from the same experiment. Thetitle compound of Example C(130) stirred in a mixture of 2.5% aq. KOH (5eq) in tetrahydrofuran for one hour. The crude product mixture wasseparated via flash column chromatography with 5% MeOH/CH₂Cl₂ to furnishthe two title compounds, as yellow amorphous solids in 30 and 50%yields, respectively, of Examples N(1) and N(2).

[1352] For Example N(1):[4-Amino-2-(4-nitro-phenylamino)-thiazol-5-yl]-(2-hydroxy-phenyl)-methanone:

[1353]¹H NMR (DMSO-d₆): δ 11.40 (1H, s), 11.00 (1H, s), 8.24 (4H, d,J=9.3 Hz), 7.89 (2H, d, J=9.3 Hz), 7.47 (1H, d, J=6.9 Hz), 7.34 (1H, dd,J=7.9, 7.7 Hz), 6.92 (2H, d, J=7.8 Hz).

[1354] HRFABMS: Calcd. for C₁₆H₁₂N₄O₄S (MH⁺): 357.0658. Found: 357.0660.

[1355] For Example N(2):N-[5-(2-Hydroxy-benzoyl)-2-(4-nitro-phenylamino)-thiazol-4-yl]-benzamide:

[1356]¹H NMR (DMSO-d₆): δ 11.80 (1H, s), 11.60 (1H, s), 10.30 (1H, s),8.27 (2H, d, J=9.2 Hz), 8.00 (2H, d, J=9.2 Hz), 7.92 (2H, d, J=7.1 Hz),7.56-7.68 (3H, m), 7.43 (1H, dd, J=7.6, 1.6 Hz), 7.34 (1H, ddd, J=8.5,7.0, 1.6 Hz), 6.94 (1H, d, J=8.2 Hz), 6.89 (1H, dd, J=7.6, 7.5 Hz).

[1357] ESIMS: Calcd. for C₂₃H₁₆N₄O₅S (MH⁺): 461. Found: 461.

[1358] Other compounds may be made in accordance with the invention inmanners similar to those described above. Additional exemplary compoundsof the invention are identified in Tables I, II, and III below, whichprovide results of biochemical and biological assays.

[1359] Biochemical and Biological Evaluation:

[1360] Cyclin-dependent kinase activity was measured by quantifying theenzyme-catalyzed, time-dependent incorporation of radioactive phosphatefrom [³²P]ATP or [³³P]ATP into a protein substrate. Unless notedotherwise, assays were performed in 96-well plates in a total volume of50 μL, in the presence of 10 mM HEPES(N-[2-hydroxyethyl]piperazine-N′-[2-ethanesulfonic acid]) (pH 7.4), 10mM MgCl₂, 25 μM adenosine triphosphate (ATP), 1 mg/mL ovalbumin, 5 μg/mLleupeptin, 1 mM dithiothreitol, 10 mM β-glycerophosphate, 0.1 mM sodiumvanadate, 1 mM sodium fluoride, 2.5 mM ethylene glycol-bis(β-aminoethylether)-N,N,N′N′-tetraacetic acid (EGTA), 2% (v/v) dimethylsulfoxide, and0.03-0.4 μCi [^(32/33)P]ATP per reaction. Reactions were initiated withenzyme, incubated at 30° C., and terminated after 20 minutes by theaddition of ethylenediaminetetraacetic acid (EDTA) to 250 mM. Thephosphorylated substrate was then captured on a nitrocellulose orphosphocellulose membrane using a 96-well filtration manifold, andunincorporated radioactivity was removed by repeated washing with 0.85%phosphoric acid. Radioactivity was quantified by exposing the driedmembranes to a phosphorimager.

[1361] Apparent K_(i) values were measured by assaying enzyme activityin the presence of different inhibitor compound concentrations andsubtracting the background radioactivity measured in the absence ofenzyme. The kinetic parameters (kcat, Km for ATP) were measured for eachenzyme under the usual assay conditions by determining the dependence ofinitial rates on ATP concentration. Inhibition data were fit to anequation for competitive inhibition using Kaleidagraph (SynergySoftware), or were fit to an equation for competitive tight-bindinginhibition using the software KineTic (BioKin, Ltd.).

[1362] Inhibition of CDK4/Cyclin D Retinoblastoma Kinase Activity:

[1363] A complex of human CDK4 and cyclin D3, or a complex of cyclin D1and a fusion protein of human CDK4 and gluathione-S-transferase(GST-CDK4), or a complex of human CDK4 and genetically truncated (1-264)cyclin D3, was purified using traditional biochemical chromatographictechniques from insect cells that had been co-infected with thecorresponding baculovirus expression vectors (see e.g., Meijer and Kim,“Chemical Inhibitors of Cyclin-Dependent Kinases,” Methods in Enzymol,.vol. 283 (1997), pp. 113-128.). The enzyme complex (5 or 50 nM) wasassayed with 0.3-0.5 μg of purified recombinant retinoblastoma proteinfragment (Rb) as a substrate. The engineered Rb fragment (residues386-928 of the native retinoblastoma protein; 62.3 kDa) contains themajority of the phosphorylation sites found in the native 106-kDaprotein, as well as a tag of six histidine residues for ease ofpurification. Phosphorylated Rb substrate was captured bymicrofiltration on a nitrocellulose membrane and quantified using aphosphorimager as described above. For measurement of tight-bindinginhibitors, the enzyme complex concentration was lowered to 5 nM, andthe assay duration was extended to 60 minutes, during which thetime-dependence of product formation was linear.

[1364] Inhibition of CDK2/Cyclin A Retinoblastoma Kinase Activity:

[1365] CDK2 was purified using published methodology (Rosenblatt et al.,“Purification and Crystallization of Human Cyclin-dependent Kinase 2,”J. Mol. Biol., vol. 230, 1993, pp. 1317-1319) from insect cells that hadbeen infected with a baculovirus expression vector. Cyclin A waspurified from E. coli cells expressing full-length recombinant cyclin A,and a truncated cyclin A construct was generated by limited proteolysisand purified as described previously (Jeffrey et al., “Mechanism of CDKactivation revealed by the structure of a cyclin A-CDK2 complex,”Nature, vol. 376 (Jul. 27, 1995), pp. 313-320). Purified, proteolyzedcyclin A was included in the assay at a three- to five-fold molar excessto CDK2. Alternatively, a complex of CDK2 and proteolyzed cyclin A wasprepared and purified by gel filtration. The substrate for this assaywas the same Rb substrate fragment used for the CDK4 assays, and themethodology of the CDK2/cyclin A and the CDK4/cyclin D3 assays wasessentially the same, except that CDK2 was present at 150 nM or 5 nM.K_(i) values were measured as described above.

[1366] Inhibition of CDK1(cdc2)/Cyclin B Histone H1 Kinase Activity:

[1367] The complex of human CDK1 (cdc2) and cyclin B was purchased fromNew England Biolabs (Beverly Mass.). Alternatively, aCDK1/glutathione-S-transferase-cyclin B1 complex was purified usingglutathione affinity chromatography from insect cells that had beenco-infected with the corresponding baculovirus expression vectors. Theassay was executed as described above at 30° C. using 2.5 units ofcdc2/cyclin B, 10 μg Histone H1 protein, and 0.1-0.3 μCi [^(32/33)P]ATPper assay. Phosphorylated histone substrate was captured bymicrofiltration on a phosphocellulose P81 membrane and quantified usinga phosphorimager as described above. K_(i) values were measured usingthe described curve-fitting programs.

[1368] Results of assays performed on compounds, which include thespecific examples described above as well as additional examplesdesignated by the prefix “I” (e.g., Examples I(1), I(2), etc.), where“*” denotes a compound having a known structure (i.e., the compound perse is known), are provided below in Tables I, II, and III. Unlessindicated otherwise in a particular entry, the units and assays used areas indicated in the applicable column of the table. The abbreviation“N.I.” indicates that no inhibition was observed at the concentrationindicated. TABLE I K_(i) with CDKs K_(i) K_(i) K_(i) CDK CDK CDK 4/D 2/A1/B Example Structure (nM) (nM) (μM) I(1)*

640^(a); 102^(b) 460 0.5 I(2)*

8000^(a);˜30 μM^(b) 8700 I(3)*

>5 μM^(a); >100 μM^(b) A(1)

660^(a); 770^(b) 1200 D(1)

490^(b) 900 I(4)*

3.1 μM^(b) 4.3 μM 3.9 I(5)*

1100^(a)870^(b) 4600 4.5 C(1)

N.I. at 10 μM^(a) N.I. at 10 μM C(2)

>5000^(a) N.I. at 100 μM C(3)

1200^(a) 4700 C(4)

95^(a) 810 0.09 A(2)

˜300^(a) D(2)

>5000^(a) A(3)

2000^(a) 6100 3.5 C(5)

140^(a) 780 0.293 C(6)

>10 μM^(a) I(6)

none @10 μM^(a) D(3)

380^(a) I(7)

˜1500^(a) I(8)

4300^(a) I(9)

2500^(a) A(4)

2000^(a) 3100 2.7 I(10)

>20 μM^(a) I(11)

520^(a) I(12)

380^(a) 4170 I(13)

2400^(a) I(14)

>50 μM^(a) I(15)

440^(a) I(16)

1800^(a) I(17)

<1000^(a) I(18)

>25 μM^(a) I(19)

1600^(a) 4800 none @100 μM A(5)

97^(a) 690 0.163 E(1)

420^(a) 320 0.03 A(6)

150^(a) 292 0.052 A(7)

310^(a) A(8)

>25 μM^(a) D(4)

800^(a) A(9)

100^(a) 230 0.053 A(10)

36^(a) 318 0.057 I(20)

none @25 μM^(a) I(21)

none @25 μM^(a) I(22)

none @25 μM^(a) none @100 μM I(23)

1500^(a) A(11)

130^(a) I(24)

N.I. at 25 μM^(a) C(7)

510^(a) C(9)

740^(a) C(10)

680^(a) C(8)

27^(a) 389 0.097 C(11)

130^(a) C(12)

27^(a) 670 A(12)

9400^(a) 4100 C(13)

51^(a) C(14)

57^(a) C(15)

57^(a) C(16)

170^(a) C(25)

1300^(a) C(24)

8^(a) 248 0.046 C(22)

67^(a) C(17)

72^(a) C(18)

12900^(a) none @10 μM C(37)

7^(a) 310 0.233 C(23)

330^(a) C(19)

15.8^(a) 277 F

40.7^(a) 350 0.2 C(20)

22^(a) 145 C(21)

117^(a) 480 A(13)

250^(a) A(14)

180^(a) C(36)

13900^(a) B

N.I. at 100 μM^(a) C(31)

N.I. at 100 μM^(a) C(32)

94^(a) C(33)

57^(a) 20 C(35)

11^(a) 23 C(34)

140^(a) 131 C(26)

330^(a) C(27)

1020^(a) C(28)

240^(a) C(29)

357^(a) C(30)

1400^(a) C(38)

25^(a) 39 C(39)

>100 μM^(a) E(2)

1170^(a) C(40)

3840^(a) C(41)

350^(a) 336 C(42)

750^(a) 207 C(43)

315^(a) C(44)

˜128^(a) C(45)

51^(a) 103 0.249 C(46)

244^(a) 1790 C(47)

30^(a) 26 C(48)

14^(a) 11.1 0.015 C(49)

23^(a) 10 0.034 C(50)

25.5^(a) 5.6 0.029 C(51)

85^(a) 33.3 C(52)

11^(a) 105 C(53)

85^(a) 180 C(54)

34^(a) 453 C(55)

8.5^(a) 493 0.504 C(56)

195^(a) 1020 C(57)

30^(a) 259 C(58)

34^(a) 306 C(59)

100^(a) 135 C(60)

56^(a) 574 I(25)

639^(a) 280 C(61)

120^(a) 1200 C(62)

72^(a) 1710 C(63)

17^(a) 200 0.133 C(64)

44000^(a) 42 μM C(65)

38000^(a) 14.3 μM C(66)

53^(a) 574 C(67)

3170^(a) 13.2 μM C(68)

>30 μM^(a) C(69)

20^(a) 638 C(70)

2.8^(a) 1120 1.37 C(71)

2^(a) 482 0.827 C(72)

13.5^(a) 169 C(73)

6.3^(a) 6.8 0.02 C(74)

4800^(a) 10.6 μM C(75)

23^(a) 1080 C(76)

2000^(a) 507 C(77)

3000^(a) None @5 μM C(78)

32^(a) 83 C(79)

54^(a) 162 C(80)

3.3^(a) 220 0.325 C(81)

12.3^(a) 872 1.24 C(82)

44.2^(a) 467 C(83)

1800^(a) 1800^(a) 32 μM C(84)

53^(a) 1040 C(85)

13^(a) 5.7 0.0022 C(86)

15^(a) 1100 1.31 C(87)

9^(a) 607 0.826 C(88)

65^(a) 305 C(89)

55^(a) 326 G

23^(a) 178 C(90)

27^(a) 771 C(91)

6.8^(a) 81 C(92)

63^(a) 9.4 C(93)

285^(a) 1040 C(94)

41^(a) 1040 1.41 C(95)

25^(a) <50 0.075 D(5)

159^(a) 233 C(96)

1.5^(a) 324 0.231 H(1)

8.2^(a) 370 0.681 H(2)

3.6^(a) 474 0.361 C(97)

8.5^(a) 392 C(98)

27^(a) 565 0.72 H(3)

2.4^(a) 405 0.472 J(1)

2.3^(a) 452 0.732 C(99)

16.4^(a) 39.5 0.04 C(100)

6.5^(a) 620 0.911 C(101)

103^(a) 300 0.32 C(102)

21^(a) 49 0.017 C(103)

110^(a) 595 C(104)

190^(a) 730 C(105)

60^(a) 1060 C(106)

134^(a) 1460 J(2)

6.4^(a) 135 0.405 C(107)

13.8^(a) 12.5 C(108)

23^(a) 6.8 0.009 C(109)

83^(a) 28 0.035 C(110)

14^(a) 260 0.104 C(111)

21^(a) 216 C(112)

23^(a) 408 C(113)

17.3^(a) 238 C(114)

21^(a) 8.5 0.028 C(115)

57^(a)55^(c) 18 0.05 J(3)

15^(a)19^(c) 572 2.0 J(4)

10.2^(a) 13.5 0.022 C(116)

121^(a) 120 0.077 J(5)

6.3^(a) 331 0.76 C(117)

10^(c) 423 0.417 C(118)

10.3^(a) 191 0.097 C(119)

24^(a) 86 0.247 C(120)

10.9^(a) 80 0.062 C(121)

10.6^(c) 953 K

43^(c) 364 C(122)

35^(c) 165 J(6)

8.1^(c) 548 0.511 C(123)

15.4^(a) 164 C(124)

17^(c) 611 0.68 C(125)

5.4^(c) 602 0.65 L

22^(c) 0.193 M

46^(c) 290 C(126)

24^(c) 390 C(127)

26^(c) 215 C(128)

30^(c) 440 C(129)

64^(c) 270

[1369] Inhibition of Cell Growth: Assessment of Cytotoxicity:

[1370] Inhibition of cell growth was measured using the tetrazolium saltassay, which is based on the ability of viable cells to reduce3-(4,5-dimethylthiazol-2-yl)-2,5-[2H]-diphenyltetrazolium bromide (MTT)to formazan (Mossman, Journal of Immunological Methods, vol. 65 (1983),pp. 55-58). The water-insoluble purple formazan product was thendetected spectrophotometrically. Various cell lines (HCT-116, Saos-2,U2-OS, SW480, COLO-205, RXF-393, M14, MDA-MB-468, and MCF7) were grownin 96-well plates. Cells were plated in the appropriate medium at avolume of 135 μl/well in either McCoy's 5A Medium (for Saos-2, U2-OS,SW480, and HCT-116 cells), RPMI (for COLO-205, RXF-393, M14 cells), orMinimum Essential Medium Eagle (for MDA-MB-468 and MCF7 cells). Plateswere incubated for four hours before addition of inhibitor compounds.Different concentrations of inhibitor compounds were added in 0.5% (v/v)dimethylsulfoxide (15 μL/well), and cells were incubated at 37° C. (5%CO₂) for four to six days (depending on cell type). At the end of theincubation, MTT was added to a final concentration of 0.2 mg/mL, andcells were incubated for 4 hours more at 37° C. After centrifugation ofthe plates and removal of medium, the absorbance of the formazan(solubilized in dimethylsulfoxide) was measured at 540 nm. Theconcentration of inhibitor compound causing 50% inhibition of growth wasdetermined from the linear portion of a semi-log plot of inhibitorconcentration versus percentage inhibition. All results were compared tocontrol cells treated only with 0.5% (v/v) dimethylsulfoxide. TABLE IIIC₅₀ with Various Cancer Cell Lines in MTT Assay IC₅₀, IC₉₀ (μM) ExampleCOLO- M14 RXF- MDA- Compound HCT116 U2-OS Saos-2 205 melanoma 393 MCF-7MB-468 SW-480 A(1) 2.5, 10 4, 9 C(4) 0.7, 30 0.4, 3 A(3) 41% @ 30 15% @30 C(5) 6, 25 3, 9 A(4) 29% @ 30 0 @ 10, 66% @ 30 I(19) 16% @ 30 22,(58% @ 30) A(5) 26, (85% @ 30) 1.7, 3 E(1) 17% @ 10 & 30 23% @ 30 A(6)20, 30 5, 15 A(9) 20, 30 4, 10 A(10) 0.95, 1.8 0.9, 1.6 1.3, 4.6 0.65,5.5 0.72, 1.7 0.7, 7.5 C(8) 18, >25 (88%) 10, >25 (84%) 6.2, 18.015, >25 6.8, 15 8.3, 20.0 (82%) C(12) 2.0, 5.0 1.9, 6.0 2.1, 4.8 C(24)0.7, 1.5 1.9, 3.0 1.4, 2.8 C(37) 10.0, 25.0 16.0, >25 (80%) 14.0, >25(70%) C(19) 9.0, 23.0 15.0, 7.0, 18.0 >25 (77%) F 1.0, 2.6 1.0, 20.01.0, 1.6 C(20) 6.1, 25 6.5, >25 (74%) 4.9, 13.0 C(21) 9.0, 22.0 22.0,13.0, 23.0 >25 (57%) C(35) 9.0, 19.0 12.0, 22.0 8.0, 20.0 C(38) 4.3,19.0 11.0, 3.0, >25 (87%) >25 (80%) C(45) 2.0, 8.0 5.5, 12.0 2.0, 10.0C(47) 4.2, 7.8 10.0, 20.0 2.5, 5.1 C(48) 0.34, 0.70 1.0, 1.5 0.42, 0.750.29, 0.60 0.6, 1.8 0.8, 1.6 2.6, >25 0.26, (82%) 0.60 C(49) 5.6, 12.014.0, 22.0 4.0, 20.0 C(50) 0.61, 1.6 2.0, 3.0 1.3, 2.8 3.0, >25 0.48,1.5 (88%) C(52) 8.5, 17.0 15.0, 23.0 9.0, 22.0 C(55) 4.3, 17.0 18, >25(74%) 17, >25 (69%) C(63) 12.0, 22.0 16.0, 9.0, 20.0 >25 (84%) C(69)5.0, 15.0 14.0, 26.0 12.0, 26.0 C(70) 2.2, 5.9 5.0, 11.0 5.8, 12.0 C(71)1.7, 4.4 3.3, 6.0 4.0, 7.0 1.2, >5.0 2.0, 1.8 >5.0 C(73) 1.4, 3.4 4.1,9.5 1.3, 5.0 C(80) 0.4, 1.1 1.0, 2.3 0.9, 1.5 0.28, 1.0 0.92, 2.5 0.84,0.4, 1.7 0.49, 1.3 1.3 C(81) 10.0, 20.0 7.9, 12.0 13.0, 22.0 >5.03.8, >5 >5.0 3.9, 16.0 4.7, 9.3 C(82) >25 (1%) >25 (1%) >25 (15%) C(85)0.25, 0.56 1.7, 2.8 0.28, 0.67 0.24, 0.58 0.71, 2.2 1.2, 3.0 3.9, 13.00.22, 0.59 C(86) 6.5, 17.0 14, 22 12, 22 C(87) 0.95, 2.9 1.8, 4.0 2.0,4.0 0.39, 1.7 1.5, 4.2 1.5, 2.3 C(88) 19, >25 (70%) >25 (26%) 22, >25(56%) C(91) 4.7, 18.0 19.0, >25 5.9, 23.0 C(94) 1.3, 3.1 3.4, 6.0 2.2,5.0 C(95) 1.5, 3.0 3.9, 5.8 1.8, 4.5 D(5) 1.9, 5.0 >25 17.0, 25.0 C(96)18.0, >25 (82%) 8.0, 12.0 4.2, 10.0 H(1) 0.58, 1.4 0.85, 1.5 0.73, 1.40.2, 0.6 2.4, 9.0 0.46, 0.88 H(2) 15.0, 25.0 16.0, 21.0 13.0, 22.0 C(97)3.9, 11.0 10.0, 21.0 9.0, 20.0 C(98) 1.3, 2.9 2.4, 5.2 2.7, 5.3 H(3)0.88, 2.4 3.5, 5.8 1.8, 3.0 J(1) 1.3, 3.0 3.5, 5.9 1.0, 5.9 C(99) 0.88,2.7 4.0, 8.0 1.1, 2.9 C(100) 2.3, 6.1 12.0, 22.0 4.5, 10.0 C(102) 1.1,2.4 3.1, 5.4 0.88, 2.2 J(2) 0.3, 0.73 1.7, 2.8 0.58, 1.3 0.9, 2.7 0.65,2.0 0.55, 0.48, 1.7 0.34, 0.7 1.1 C(107) 2.5, 7.0 7.9, 12.0 5.7, 12.0C(108) 0.4, 1.4 1.4, 4.8 0.31, 3.2 C(109) 1.6, 3.0 4.7, 19.0 1.5, 14.0C(110) 0.64, 1.7 0.7, 1.6 0.89, 1.8 0.4, 1.7 0.44, 0.75 C(111) 1.7, 3.83.0, 5.5 3.8, 5.9 C(113) 3.8, 8.0 8.0, 19.0 2.8, 9.0 C(114) 1.1, 2.83.0, 5.2 1.3, 4.8 C(115) 0.98, 2.1 2.7, 5.0 0.8, 2.2 J(3) 0.9, 2.8 2.9,5.2 2.2, 4.9 J(5) 0.7, 1.6 1.0, 1.8 0.9, 1.5 C(117) 1.8, 2.9 0.9, 1.70.89, 1.5 C(118) 0.64, 1.4 2.4, 5.0 0.94, 1.5 C(119) 4.0, 7.0 7.8, 12.07.0, 13.0 C(120) 3.5, 6.0 5.7, 11.0 2.6, 5.2 C(121) 2.5, 5.3 6.0, 11.05.1, 12.0 K 3.5, 6.2 3.5, 6.0 6.0, 13.0 C(122) 1.7, 4.8 1.5, 4.7 3.7,10.0 J(6) 0.26, 0.6 0.51, 1.3 0.47, 0.77 C(123) 2.7, 7.0 7.1, 12.0 2.9,5.7 C(124) 0.54, 1.6 1.3, 2.5 0.98, 1.6 C(125) 0.62, 1.8 1.2, 2.3 0.9,1.5 L >12.5 M >25 C(126) 0.6, 1.4

[1371] pRb Immunoblotting:

[1372] The ability of compounds to inhibit phosphorylation of theretinoblastoma protein (pRb) was assessed by western blot analysis. Ananti-Rb antibody was used to measure the conversion ofhyper-phosphorylated pRb to hypo-phosphorylated pRb. An anti-phospho-Rb(ser780) antibody was used to specifically measure dephosphorylation atserine 780, a site that has previously been shown to be phosphorylatedby CDK4/cyclin D. Inhibition of pRb phosphorylation is indicated by a“+” in Table III below, and failure to inhibit pRb phosphorylation isindicated by a “−” in the table.

[1373] Human colon tumor cells (HCT-116 cells; 5×10⁶) were plated on 100mM dishes and allowed to grow overnight. Five micromolar of eachcompound was added for 12 hours. The cells were then collected andcentrifuged. The cell pellets were lysed by the addition of 100 μL lysisbuffer (50 mM HEPES (pH 7.0), 250 mM NaCl, 5 mMethylenediaminetetraacetic acid, 0.1% Nonidet P-40, 1 mM dithiothreitol,2 mM sodium pyrophosphate, 1 mM sodium orthovanadate, 1 μg/ml aprotonin,1 μg/ml leupeptin, 50 μg/ml phenylmethylsulfonyl fluoride). Fortymicrograms of protein were separated by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) on a 6% gel. Theproteins were transferred to nitrocellulose and blocked with 5% blockingbuffer in Tris-buffered saline overnight. The anti-Rb antibody(Pharmingen), the anti-phospho-Rb (Ser 780) antibody (MBL), andsecondary antibody were incubated for 1 hour at room temperaturefollowed by three wash steps in 0.01% Tween-20 in Tris-buffered saline.The Rb protein was detected using chemiluminescence according to themanufacturer (Amersham). TABLE III Inhibition of pRb PhosphorylationInhibits Inhibits pRb pRb (ser 780) Example phosphoryl- phosphoryl-Compound ation ation C(85) + + J(2) + + C(80) + + C(48) + + H(1) + +C(50) + + C(87) + + C(73) + + C(81) + + C(94) + + F − − C(71) + +

[1374] The examples above illustrate compounds according to Formula Iand assays that may readily be performed to determine their activitylevels against the various CDK/cyclin complexes. It will be apparentthat such assays or other suitable assays known in the art may be usedto select an inhibitor having a desired level of activity against aselected target.

[1375] While the invention has been illustrated by reference to specificand preferred embodiments, those skilled in the art will recognize thatvariations and modifications may be made through routine experimentationand practice of the invention. For example, those of ordinary skill inthe art will recognize that variations or substitutions to the compoundsof Formula I may be made without adversely affecting in a significantmanner their efficacy in the pharmaceutical compositions. Thus, theinvention is intended not to be limited by the foregoing description,but to be defined by the appended claims and their equivalents.

What is claimed is:
 1. A compound of the Formula I:

wherein: R¹ is a substituted or unsubstituted group selected from:C₁₋₆-alkyl; C₁₋₆-alkenyl; C₁₋₆-alkynyl; C₁₋₆-alkoxyl; C₁₋₆-alcohol;carbocyclic or heterocyclic, monocyclic or fused or non-fusedpolycyclic, cycloalkyl; carbocyclic or heterocyclic, monocyclic or fusedor non-fused polycyclic, aryl; carbonyl; ether; (C₁₋₆-alkyl)-carbonyl;(C₁₋₆-alkyl)-aryl; (C₁₋₆-alkyl)-cycloalkyl; (C₁₋₆-alkyl)-(C₁₋₆-alkoxyl);aryl-(C₁₋₆-alkoxyl); thioether; thiol; and sulfonyl; wherein when R¹ issubstituted, each substituent independently is a halogen; haloalkyl;C₁₋₆-alkyl; C₁₋₆-alkenyl; C₁₋₆-alkynyl; hydroxyl; C₁₋₆-alkoxyl; amino;nitro; thiol; thioether; imine; cyano; amido; phosphonato; phosphine;carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; ester;oxygen; carbocyclic or heterocyclic, monocyclic or fused or non-fusedpolycyclic, cycloalkyl; or carbocyclic or heterocyclic, monocyclic orfused or non-fused polycyclic, aryl; and R² is a carbocyclic orheterocyclic, monocyclic or fused or non-fused polycyclic, ringstructure having a substituent at the position adjacent to the point ofattachment, which ring structure is optionally further substituted,where each substituent of R² independently is a halogen; haloalkyl;C₁₋₆-alkyl; C₁₋₆-alkenyl; C₁₋₆-alkynyl; hydroxyl; C₁₋₆-alkoxyl; amino;nitro; thiol; thioether; imine; cyano; amido; phosphonato; phosphine;carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; ester;oxygen; carbocyclic or heterocyclic, monocyclic or fused or non-fusedpolycyclic, cycloalkyl; or carbocyclic or heterocyclic, monocyclic orfused or non-fused polycyclic, aryl; or a pharmaceutically acceptablesalt of a compound of the Formula I, or a prodrug or pharmaceuticallyactive metabolite of a compound of the Formula I or pharmaceuticallyacceptable salt thereof.
 2. A compound, pharmaceutically acceptablesalt, prodrug, or pharmaceutically active metabolite thereof accordingto claim 1, wherein: when R¹ is substituted, each substituentindependently is a halogen, haloalkyl, C₁₋₆-alkyl, C₁₋₆-alkenyl,C₁₋₆-alkynyl, hydroxyl, oxygen, C₁₋₆-alkoxyl, amino, nitro, thiol,thioether, imine, cyano, amido, phosphonato, phosphine, carboxyl,thiocarbonyl, sulfonyl, sulfonamide, ketone, aldehyde, or ester; andeach substituent of R² independently is a halogen, haloalkyl,C₁₋₆-alkyl, C₁₋₆-alkenyl, C₁₋₆-alkynyl, hydroxyl, C₁₋₆-alkoxyl, amino,nitro, thiol, thioether, imine, cyano, amido, phosphonato, phosphine,carboxyl, thiocarbonyl, sulfonyl, sulfonamide, ketone, aldehyde, orester.
 3. A compound, pharmaceutically acceptable salt, prodrug, oractive metabolite according to claim 1, wherein R¹ is a substitutedphenyl group.
 4. A compound, pharmaceutically acceptable salt, prodrug,or active metabolite according to claim 1, wherein R¹ is phenylsubstituted with an alkylamine or pyridine group.
 5. A compound,pharmaceutically acceptable salt, prodrug, or active metaboliteaccording to claim 1, wherein R¹ is selected from the group consistingof:


6. A compound, pharmaceutically acceptable salt, prodrug, or activemetabolite according to claim 1, wherein R¹ is phenyl substituted byoptionally substituted carbonyl or sulfonamide.
 7. A compound,pharmaceutically acceptable salt, prodrug, or active metaboliteaccording to claim 1, wherein R¹ is selected from the group consistingof:

where R³ is selected from the group consisting of C₁-C₆ alkyl, C₁-C₆alkoxy, aryl, aryloxy, and amine.
 8. A compound, pharmaceuticallyacceptable salt, prodrug or active metabolite according to claim 1,wherein R² is ortho-substituted phenyl or thienyl.
 9. A compound,pharmaceutically acceptable salt, prodrug or active metabolite accordingto claim 9, wherein R² is o-halophenyl or o-dihalophenyl.
 10. Acompound, pharmaceutically acceptable salt, prodrug or active metaboliteaccording to claim 10, wherein R² is o-difluorophenyl.
 11. A compoundaccording to claim 1 selected from the group consisting of:

or a pharmaceutically acceptable salt, prodrug, or active metabolite ofsaid compound.
 12. A compound according to claim 1 selected from thegroup consisting of:

or a pharmaceutically acceptable salt, prodrug, or active metabolite ofsaid compound.
 13. A compound selected from the group consisting of:

or a pharmaceutically acceptable salt, prodrug, or active metabolite ofsaid compound.
 14. A pharmaceutical composition comprising: (a) anamount of a cell-cycle control agent effective to inhibit CDK4 or aCDK4/cyclin complex, said cell-cycle control agent being selected fromthe group consisting of: (i) a compound of the Formula I:

 wherein: R¹ is a substituted or unsubstituted group selected from:C₁₋₆-alkyl; C₁₋₆-alkenyl; C₁₋₆-alkynyl; C₁₋₆-alkoxyl; C₁₋₆-alcohol;carbocyclic or heterocyclic, monocyclic or fused or non-fusedpolycyclic, cycloalkyl; carbocyclic or heterocyclic, monocyclic or fusedor non-fused polycyclic, aryl; carbonyl; ether; (C₁₋₆-alkyl)-carbonyl;(C₁₋₆-alkyl)-aryl; (C₁₋₆-alkyl)-cycloalkyl; (C₁₋₆-alkyl)-(C₁₋₆-alkoxyl);aryl-(C₁₋₆-alkoxyl); thioether; thiol; and sulfonyl; wherein when R¹ issubstituted, each substituent independently is a halogen; haloalkyl;C₁₋₆-alkyl; C₁₋₆-alkenyl; C₁₋₆-alkynyl; hydroxyl; C₁₋₆-alkoxyl; amino;nitro; thiol; thioether; imine; cyano; amido; phosphonato; phosphine;carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; ester;oxygen; carbocyclic or heterocyclic, monocyclic or fused or non-fusedpolycyclic, cycloalkyl; or carbocyclic or heterocyclic, monocyclic orfused or non-fused polycyclic, aryl; and R² is a carbocyclic orheterocyclic, monocyclic or fused or non-fused polycyclic, ringstructure having a substituent at the position adjacent to the point ofattachment, which ring structure is optionally further substituted,where each substituent of R² independently is a halogen; haloalkyl;C₁₋₆-alkyl; C₁₋₆-alkenyl; C₁₋₆-alkynyl; hydroxyl; C₁₋₆-alkoxyl; amino;nitro; thiol; thioether; imine; cyano; amido; phosphonato; phosphine;carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; ester;oxygen; carbocyclic or heterocyclic, monocyclic or fused or non-fusedpolycyclic, cycloalkyl; or carbocyclic or heterocyclic, monocyclic orfused or non-fused polycyclic, aryl; (ii) a pharmaceutically acceptablesalt of a compound of the Formula I; and (iii) a prodrug orpharmaceutically active metabolite of a compound of the Formula I or apharmaceutically acceptable salt thereof; and (b) a pharmaceuticallyacceptable carrier.
 15. A method of treating a disease or disordermediated by inhibition of CDK4 or a CDK4/cyclin complex, comprisingadministering to a subject in need of such treatment a cell-cyclecontrol agent selected from the group consisting of: compounds of theFormula I:

 wherein: R¹ is a substituted or unsubstituted group selected from:C₁₋₆-alkyl; C₁₋₆-alkenyl; C₁₋₆-alkynyl; C₁₋₆-alkoxyl; C₁₋₆-alcohol;carbocyclic or heterocyclic, monocyclic or fused or non-fusedpolycyclic, cycloalkyl; carbocyclic or heterocyclic, monocyclic or fusedor non-fused polycyclic, aryl; carbonyl; ether; (C₁₋₆-alkyl)-carbonyl;(C₁₋₆-alkyl)-aryl; (C₁₋₆-alkyl)-cycloalkyl; (C₁₋₆-alkyl)-(C₁₋₆-alkoxyl);aryl-(C₁₋₆-alkoxyl); thioether; thiol; and sulfonyl; wherein when R¹ issubstituted, each substituent independently is a halogen; haloalkyl;C₁₋₆-alkyl; C₁₋₆-alkenyl; C₁₋₆-alkynyl; hydroxyl; oxygen; C₁₋₆-alkoxyl;amino; nitro; thiol; thioether; imine; cyano; amido; phosphonato;phosphine; carboxyl; thiocarbonyl; sulfonyl; sulfonamide; ketone;aldehyde; or ester; and R² is a carbocyclic or heterocyclic, monocyclicor fused or non-fused polycyclic, ring structure having a substituent atthe position adjacent the point of attachment, which ring structure isoptionally further substituted, where each substituent of R²independently is a halogen; haloalkyl; C₁₋₆-alkyl; C₁₋₆-alkenyl;C₁₋₆-alkynyl; hydroxyl; C₁₋₆-alkoxyl; amino; nitro; thiol; thioether;imine; cyano; amido; phosphonato; phosphine; carboxyl; thiocarbonyl;sulfonyl; sulfonamide; ketone; aldehyde; or ester; pharmaceuticallyacceptable salts of compounds of the Formula I; and prodrugs andpharmaceutically active metabolites of compounds of the Formula I andtheir pharmaceutically acceptable salts.
 16. A compound of the FormulaI:

wherein: R¹ is selected from:

R² is a substituted or unsubstituted: carbocyclic or heterocyclic,monocyclic or fused or non-fused polycyclic, ring structure; where eachoptional substituent for R² is independently a halogen; oxygen;haloalkyl; C₁₋₆-alkyl; C₁₋₆-alkenyl; C₁₋₆-alkynyl; hydroxyl;C₁₋₆-alkoxyl; carbocyclic or heterocyclic, monocyclic or fused ornon-fused polycyclic, cycloalkyl; carbocyclic or heterocyclic,monocyclic or fused or non-fused polycyclic, aryl; amino; nitro; thiol;thioether; imine; cyano; amido; phosphonato; phosphine; carboxyl;thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; or ester; or apharmaceutically acceptable salt of a compound of the Formula I, or aprodrug or pharmaceutically active metabolite of a compound of theFormula I or pharmaceutically acceptable salt thereof.
 17. Apharmaceutical composition comprising: (a) an effective amount forinhibiting a CDK or a CDK/cyclin complex of a cell-cycle control agentselected from: (i) compounds of the Formula I:

 wherein: R¹ is selected from:

R² is a substituted or unsubstituted: carbocyclic or heterocyclic,monocyclic or fused or non-fused polycyclic, ring structure; where eachoptional substituent for R² is independently a halogen; oxygen;haloalkyl; C₁₋₆-alkyl; C₁₋₆-alkenyl; C₁₋₆-alkynyl; hydroxyl;C₁₋₆-alkoxyl; carbocyclic or heterocyclic, monocyclic or fused ornon-fused polycyclic, cycloalkyl; carbocyclic or heterocyclic,monocyclic or fused or non-fused polycyclic, aryl; amino; nitro; thiol;thioether; imine; cyano; amido; phosphonato; phosphine; carboxyl;thiocarbonyl; sulfonyl; sulfonamide; ketone; aldehyde; or ester; (ii)pharmaceutically acceptable salts of compounds of the Formula I; and(iii) prodrugs and pharmaceutically active metabolites of compounds ofthe Formula I or pharmaceutically acceptable salts thereof; and (b) apharmaceutically acceptable carrier.